IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation human-b-cell Homo sapiens B cell b-cell B-cell [B lymphocyte; B-lymphocyte; B cell; B-cells] pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. Naïve B cells isolated from peripheral blood. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation mouse-spleen Mus musculus spleen spleen spleen pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. B1-8 transgenic mice harboring NIP-specific B cells. "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. FACS after "1-PLA". NIP-coupled latex beads were bound to purified soluble forms of monomeric or pentameric IgM. "1-PLA":- coupling of the DNA oligos directly to primary antibody. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation mouse-tko-md-pre-b cells Mouse TKO_MD Pre B Cells:KO for SLP-65, RAG2, CD179b, transfected with NOP specific IgM/IgD BCR tko_md_pre-b-cells TKO_MD Pre B Cells:KO for SLP-65, RAG2, CD179b, transfected with NOP specific IgM/IgD BCR Transfected with IgM- and IgD-BCR specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP) pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. "1-PLA":- coupling of the DNA oligos directly to primary antibody. TKO-MD is a pro-B cell line derived from RAG, Lambda5, SLP-65 triple-deficient mouse bone marrow which is transfected to express an IgM- and IgD-BCR specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP). All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation mouse-tko-md-pre-b cells Mouse TKO_MD Pre B Cells:KO for SLP-65, RAG2, CD179b, transfected with NOP specific IgM/IgD BCR tko_md_pre-b-cells TKO_MD Pre B Cells:KO for SLP-65, RAG2, CD179b, transfected with NOP specific IgM/IgD BCR Transfected with IgM- and IgD-BCR specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP) pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. TKO-MD is a pro-B cell line derived from RAG, Lambda5, SLP-65 triple-deficient mouse bone marrow which is transfected to express an IgM- and IgD-BCR specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP). "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation in vitro In vitro facs fluorescence-activated cell sorting FACS Flow cytometry facs predetermined predetermined participant predetermined Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. FACS after Fab-PLA. NIP-coupled latex beads were bound to purified soluble forms of monomeric or pentameric IgM. "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation in vitro In vitro facs fluorescence-activated cell sorting FACS Flow cytometry facs predetermined predetermined participant predetermined Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. FACS after "1-PLA". NIP-coupled latex beads were bound to purified soluble forms of monomeric or pentameric IgM. "1-PLA":- coupling of the DNA oligos directly to primary antibody. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation mouse-tko-pre-b-cells Mouse Triple_ Knock_out pre-B cells:SLP-65,Rag2, CD179b deficient tko_pre-b cells Triple_ Knock_out pre-B cells:SLP-65,Rag2, CD179b deficient pla proximity ligation assay pELISA pELISA PLA autoradiography autoradiography Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. TKO-D is a pro-B cell line derived from RAG, Lambda5, SLP-65 triple-deficient mouse bone marrow which is transfected to express an IgD specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP). "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation mouse-tko-md-pre-b cells Mouse TKO_MD Pre B Cells:KO for SLP-65, RAG2, CD179b, transfected with NOP specific IgM/IgD BCR tko_md_pre-b-cells TKO_MD Pre B Cells:KO for SLP-65, RAG2, CD179b, transfected with NOP specific IgM/IgD BCR Transfected with IgM- and IgD-BCR specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP) pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. TKO-MD is a pro-B cell line derived from RAG, Lambda5, SLP-65 triple-deficient mouse bone marrow which is transfected to express an IgM- and IgD-BCR specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP). "2-PLA":- usual PLA protocol;secondary antibodies linked to oligo-probes targetted to primary antibody. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation mouse-tko-pre-b-cells Mouse Triple_ Knock_out pre-B cells:SLP-65,Rag2, CD179b deficient tko_pre-b cells Triple_ Knock_out pre-B cells:SLP-65,Rag2, CD179b deficient pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. TKO-M is a pro-B cell line derived from RAG, Lambda5, SLP-65 triple-deficient mouse bone marrow which is transfected to express an IgM specific for the hapten 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP). "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 2014 imex curation Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Only protein-protein interactions imex curation drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kläsener K., Maity P.C., Hobeika E., Yang J., Reth M. Affinomics - Interactions curated for the Affinomics consortium. Interactions shown in Supp fig 2 cannot be annotated as the identity of the Lambda LC is not known; the antibody used (JC5-1 clone) detects lambda 1 and lambda 2 light chains. Some interactions from Figure 3 (using the mutant) and figure 6 cannot be annotated as the construct origins or identities are not known. In Figure 6 artificial constructs are also used; results obtained from their use are of dubious biological relevence. I regret starting this; as it is a general pain, and not much useful annotation for a lot of work... email sent regarding species in figure 3 (already entered as Mus) and Fig 6 - - no reply. "FAB-PLA":-coupling of the DNA oligos directly to Fab fragments. All anti-IgM antibodies are targetted to the heavy chain (constant region). IgD:- It has been assumed the constant region of the IgD heavy chain has also been targetted ;however the immunogen is not stated for some of the possible antibodies used. michael.reth@bioss.uni-freiburg.de Accepted 2015-APR-07 by ORCHARD Elife 2014 Only protein-protein interactions imex curation imex curation I have altered EBI-10683395. For Fig 2B I have only added the pentamer, as the presence of a monomer is not an "interaction". cd19_mouse B-lymphocyte antigen CD19 Differentiation antigen CD19 Cd19 protein protein mouse Mus musculus Mouse MPSPLPVSFLLFLTLVGGRPQKSLLVEVEEGGNVVLPCLPDSSPVSSEKLAWYRGNQSTPFLELSPGSPGLGLHVGSLGILLVIVNVSDHMGGFYLCQKRPPFKDIWQPAWTVNVEDSGEMFRWNASDVRDLDCDLRNRSSGSHRSTSGSQLYVWAKDHPKVWGTKPVCAPRGSSLNQSLINQDLTVAPGSTLWLSCGVPPVPVAKGSISWTHVHPRRPNVSLLSLSLGGEHPVREMWVWGSLLLLPQATALDEGTYYCLRGNLTIERHVKVIARSAVWLWLLRTGGWIVPVVTLVYVIFCMVSLVAFLYCQRAFILRRKRKRMTDPARRFFKVTPPSGNGTQNQYGNVLSLPTSTSGQAHAQRWAAGLGSVPGSYGNPRIQVQDTGAQSHETGLEEEGEAYEEPDSEEGSEFYENDSNLGQDQVSQDGSGYENPEDEPMGPEEEDSFSNAESYENADEELAQPVGRMMDFLSPHGSAWDPSREASSLGSQSYEDMRGILYAAPQLHSIQSGPSHEEDADSYENMDKSDDLEPAWEGEGHMGTWGTT DDC34A3E23410C8D cd52_mouse CAMPATH-1 antigen Cd52 Cdw52 Mb7 Lymphocyte differentiation antigen B7 protein protein mouse Mus musculus Mouse MKSFLLFLTIILLVVIQIQTGSLGQATTAASGTNKNSTSTKKTPLKSGASSIIDAGACSFLFFANTLMCLFYLS E106F479AC481925 chtb_vibch Cholera enterotoxin subunit B ctxB toxB VC_1456 Cholera enterotoxin B chain Cholera enterotoxin gamma chain Choleragenoid protein protein vibch Vibrio cholerae MIKLKFGVFFTVLLSSAYAHGTPQNITDLCAEYHNTQIYTLNDKIFSYTESLAGKREMAIITFKNGAIFQVEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVEKLCVWNNKTPHAIAAISMAN 9AA393E3EA8E3EBF ighm_human Immunoglobulin heavy constant mu Ig mu chain C region BOT IGHM Ig mu chain C region Ig mu chain C region GAL Ig mu chain C region OU protein protein human Homo sapiens Human GSASAPTLFPLVSCENSPSDTSSVAVGCLAQDFLPDSITFSWKYKNNSDISSTRGFPSVLRGGKYAATSQVLLPSKDVMQGTDEHVVCKVQHPNGNKEKNVPLPVIAELPPKVSVFVPPRDGFFGNPRKSKLICQATGFSPRQIQVSWLREGKQVGSGVTTDQVQAEAKESGPTTYKVTSTLTIKESDWLGQSMFTCRVDHRGLTFQQNASSMCVPDQDTAIRVFAIPPSFASIFLTKSTKLTCLVTDLTTYDSVTISWTRQNGEAVKTHTNISESHPNATFSAVGEASICEDDWNSGERFTCTVTHTDLPSPLKQTISRPKGVALHRPDVYLLPPAREQLNLRESATITCLVTGFSPADVFVQWMQRGQPLSPEKYVTSAPMPEPQAPGRYFAHSILTVSEEEWNTGETYTCVVAHEALPNRVTERTVDKSTGKPTLYNVSLVMSDTAGTCY 607CA2FA7BFA0D39 cd19_human B-lymphocyte antigen CD19 CD19 Differentiation antigen CD19 B-lymphocyte surface antigen B4 T-cell surface antigen Leu-12 protein protein human Homo sapiens Human MPPPRLLFFLLFLTPMEVRPEEPLVVKVEEGDNAVLQCLKGTSDGPTQQLTWSRESPLKPFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAGKYYCHRGNLTMSFHLEITARPVLWHWLLRTGGWKVSAVTLAYLIFCLCSLVGILHLQRALVLRRKRKRMTDPTRRFFKVTPPPGSGPQNQYGNVLSLPTPTSGLGRAQRWAAGLGGTAPSYGNPSSDVQADGALGSRSPPGVGPEEEEGEGYEEPDSEEDSEFYENDSNLGQDQLSQDGSGYENPEDEPLGPEDEDSFSNAESYENEDEELTQPVARTMDFLSPHGSAWDPSREATSLGSQSYEDMRGILYAAPQLRSIRGQPGPNHEEDADSYENMDNPDGPDPAWGGGGRMGTWSTR A0E08DD628B69E51 ighd_human Immunoglobulin heavy constant delta IGHD Ig delta chain C region Ig delta chain C region NIG-65 Ig delta chain C region WAH protein protein human Homo sapiens Human APTKAPDVFPIISGCRHPKDNSPVVLACLITGYHPTSVTVTWYMGTQSQPQRTFPEIQRRDSYYMTSSQLSTPLQQWRQGEYKCVVQHTASKSKKEIFRWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLRDKATFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQHSRLTLPRSLWNAGTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAASWLLCEVSGFSPPNILLMWLEDQREVNTSGFAPARPPPQPRSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRTLLNASRSLEVSYVTDHGPMK 726F08E73519DDDC cd79a_mouse B-cell antigen receptor complex-associated protein alpha chain Cd79a Iga Mb-1 Ig-alpha MB-1 membrane glycoprotein Membrane-bound immunoglobulin-associated protein Surface IgM-associated protein protein protein mouse Mus musculus Mouse MPGGLEALRALPLLLFLSYACLGPGCQALRVEGGPPSLTVNLGEEARLTCENNGRNPNITWWFSLQSNITWPPVPLGPGQGTTGQLFFPEVNKNHRGLYWCQVIENNILKRSCGTYLRVRNPVPRPFLDMGEGTKNRIITAEGIILLFCAVVPGTLLLFRKRWQNEKFGVDMPDDYEDENLYEGLNLDDCSMYEDISRGLQGTYQDVGNLHIGDAQLEKP DIP protein P11911 original sequence version: 147 A4C648C2BE6D3E38 ighm_mouse Immunoglobulin heavy constant mu Ighm Igh-6 protein protein mouse Mus musculus Mouse SQSFPNVFPLVSCESPLSDKNLVAMGCLARDFLPSTISFTWNYQNNTEVIQGIRTFPTLRTGGKYLATSQVLLSPKSILEGSDEYLVCKIHYGGKNRDLHVPIPAVAEMNPNVNVFVPPRDGFSGPAPRKSKLICEATNFTPKPITVSWLKDGKLVESGFTTDPVTIENKGSTPQTYKVISTLTISEIDWLNLNVYTCRVDHRGLTFLKNVSSTCAASPSTDILTFTIPPSFADIFLSKSANLTCLVSNLATYETLNISWASQSGEPLETKIKIMESHPNGTFSAKGVASVCVEDWNNRKEFVCTVTHRDLPSPQKKFISKPNEVHKHPPAVYLLPPAREQLNLRESATVTCLVKGFSPADISVQWLQRGQLLPQEKYVTSAPMPEPGAPGFYFTHSILTVTEEEWNSGETYTCVVGHEALPHLVTERTVDKSTGKPTLYNVSLIMSDTGGTCY EBCCA6E8569AEEC5 cd20_mouse B-lymphocyte antigen CD20 Ms4a1 Cd20 Ly-44 Ms4a2 B-cell differentiation antigen Ly-44 Lymphocyte antigen 44 Membrane-spanning 4-domains subfamily A member 1 protein protein mouse Mus musculus Mouse MSGPFPAEPTKGPLAMQPAPKVNLKRTSSLVGPTQSFFMRESKALGAVQIMNGLFHITLGGLLMIPTGVFAPICLSVWYPLWGGIMYIISGSLLAAAAEKTSRKSLVKAKVIMSSLSLFAAISGIILSIMDILNMTLSHFLKMRRLELIQTSKPYVDIYDCEPSNSSEKNSPSTQYCNSIQSVFLGILSAMLISAFFQKLVTAGIVENEWKRMCTRSKSNVVLLSAGEKNEQTIKMKEEIIELSGVSSQPKNEEEIEIIPVQEEEEEEAEINFPAPPQEQESLPVENEIAP DF478ECD2C5C16FC cd20_human B-lymphocyte antigen CD20 MS4A1 CD20 B-lymphocyte surface antigen B1 Bp35 Leukocyte surface antigen Leu-16 Membrane-spanning 4-domains subfamily A member 1 protein protein human Homo sapiens Human MTTPRNSVNGTFPAEPMKGPIAMQSGPKPLFRRMSSLVGPTQSFFMRESKTLGAVQIMNGLFHIALGGLLMIPAGIYAPICVTVWYPLWGGIMYIISGSLLAATEKNSRKCLVKGKMIMNSLSLFAAISGMILSIMDILNIKISHFLKMESLNFIRAHTPYINIYNCEPANPSEKNSPSTQYCYSIQSLFLGILSVMLIFAFFQELVIAGIVENEWKRTCSRPKSNIVLLSAEEKKEQTIEIKEEVVGLTETSSQPKNEEDIEIIPIQEEEEEETETNFPEPPQDQESSPIENDSSP AC5420F8B626BDD1 ighdm_mouse Ig delta chain C region membrane-bound form protein protein mouse Mus musculus Mouse DKKEPDMFLLSECKAPEENEKINLGCLVIGSQPLKISWEPKKSSIVEHVFPSEMRNGNYTMVLQVTVLASELNLNHTCTINKPKRKEKPFKFPESWDSQSSKRVTPTLQAKNHSTEATKAITTKKDIEGAMAPSNLTVNILTTSTHPEMSSWLLCEVSGFFPENIHLMWLGVHSKMKSTNFVTANPTAQPGGTFQTWSVLRLPVALSSSLDTYTCVVEHEASKTKLNASKSLAISGIVNTIQHSCIMDEQSDSYMDLEEENGLWPTMCTFVALFLLTLLYSGFVTFIKVK 037A888CEB7598A0 ksyk_mouse Tyrosine-protein kinase SYK Syk Spleen tyrosine kinase Sykb ptk72 protein protein mouse Mus musculus Mouse MAGSAVDSANHLTYFFGNITREEAEDYLVQGGMTDGLYLLRQSRNYLGGFALSVAHNRKAHHYTIERELNGTYAISGGRAHASPADLCHYHSQEPDGLICLLKKPFNRPPGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGNISRDESEQTVLIGSKTNGKFLIRARDNSGSYALCLLHEGKVLHYRIDRDKTGKLSIPEGKKFDTLWQLVEHYSYKPDGLLRVLTVPCQKIGAQMGHPGSPNAHPVTWSPGGIISRIKSYSFPKPGHKKPAPPQGSRPESTVSFNPYEPTGGPWGPDRGLQREALPMDTEVYESPYADPEEIRPKEVYLDRSLLTLEDNELGSGNFGTVKKGYYQMKKVVKTVAVKILKNEANDPALKDELLAEANVMQQLDNPYIVRMIGICEAESWMLVMEMAELGPLNKYLQQNRHIKDKNIIELVHQVSMGMKYLEESNFVHRDLAARNVLLVTQHYAKISDFGLSKALRADENYYKAQTHGKWPVKWYAPECINYYKFSSKSDVWSFGVLMWEAFSYGQKPYRGMKGSEVTAMLEKGERMGCPAGCPREMYDLMNLCWTYDVENRPGFTAVELRLRNYYYDVVN DIP protein P48025 original sequence version: 108 DIP protein P48025 original sequence version: 125 2F98D21601F8224E ighm-ighm-6 4 19 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 1E Results did not distinguish between resting and antigen (NIP) stimulated cells. ighm-ighm-7 9 19 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 1D Results did not distinguish between resting and antigen (NIP) stimulated cells. ighm-ighm-2 5 19 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 1F/H, 4B The Fab-PLA signal is lost upon exposure of B cells to antigen (NIP), or in presence of BCR-activating drugs. Only Fab-PLA showed a clear difference in the relative IgM:IgM conformation or distance between resting and activated B cells. Specific antigen- NIP, pervanadate and Lat-A. ighm-ighm-8 6 19 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line physical association physical association 2C-2 Fab-PLA detects the pentameric IgM but not the loosely spaced monomeric IgM. Fab-PLA specifically detects oligomeric forms of IgM be it the soluble IgM pentamer or the oligomeric IgM-BCR on the B cell surface. ighm-1 7 19 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line 19 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line physical association physical association 2C Two IgM monomers interact. ighm-ighm-1 7 19 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line 19 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line physical association physical association 2C-2 1-PLA detects both soluble forms of IgM equally well - monomers and pentamers. ighm-ighm-9 2 19 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 4A/C, 5A/B/C, Supp 5 The Fab-PLA signal is lost upon exposure of B cells to antigen, or in presence of BCR-activating drugs. specific antigen-NIP, pervanadate (Perv) and Lat-A - activators of the BCR. When cells were pretreated with R406 - a Syk inhibitor, Lat-A activation did not occur and no loss of signal was seen. Pre-treatment with Src-family kinase inhibitor PP2 only delays the loss of signal. R406 - a Syk inhibitor, stops the loss of signal after the specific antigen-NIP stimulation. ighm-ighm-10 1 15 unspecified role unspecified role neutral component neutral component 15 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 4D Pervanadate, Lat A;BCR-activating drugs cd79a-ighm-1 2 23 unspecified role unspecified role neutral component neutral component 18 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 5D pervanadate (Perv) - activator of the BCR. When permeabilised cells were pretreated with R406 - a Syk inhibitor, prior to LatA treatment, the interaction is inhibited. Pre-treatment with Src-family kinase inhibitor PP2 only delays a positive response. Lat-A - activator of the BCR. ighdm-ighdm-3 11 22 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 3B No oligomerisation of IgD was seen when an IgD mutant was transfected. (exact mutations are not stated.) ighdm-ighdm-2 2 22 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 4A/C The Fab-PLA signal is lost upon exposure of B cells to antigen, or in presence of BCR-activating drugs. specific antigen-NIP, pervanadate (Perv) and Lat-A - activators of the BCR. ighdm-ighm-1 5 22 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 4B The Fab-PLA signal is lost upon exposure of B cells to antigen, or in presence of BCR-activating drugs. specific antigen-NIP, pervanadate (Perv) and Lat-A - activators of the BCR. ighd-ighd-1 1 17 unspecified role unspecified role neutral component neutral component 17 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 4D The Fab-PLA signal is lost upon exposure of B cells to antigen, or in presence of BCR-activating drugs. specific antigen-NIP, pervanadate (Perv) and Lat-A - activators of the BCR. cd19-ighm-1 5 19 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7A CD19 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP cd20-ighm-2 5 20 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7A-2 CD20 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP ighm-ms4a1-2 1 15 unspecified role unspecified role neutral component neutral component 21 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7C-2 CD20 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP cd19-ighm-3 1 16 unspecified role unspecified role neutral component neutral component 15 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7C CD19 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP cd19-ighm-4 5 12 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7B CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP igdhm-cd20-1 5 22 unspecified role unspecified role neutral component neutral component 20 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7B-2 CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP ighdm-ms4a1-2 1 17 unspecified role unspecified role neutral component neutral component 21 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7D-2 CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP cd19-ighdm-3 1 16 unspecified role unspecified role neutral component neutral component 17 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7D CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP cd19-ighm-5 2 19 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7E CD19 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP ighm-ms4a1-1 2 19 unspecified role unspecified role neutral component neutral component 20 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7E-2 CD20 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP cd19-ighdm-1 2 22 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7F CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP ighdm-ms4a1-1 2 20 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7F-2 CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP cd19-ighm-6 2 19 unspecified role unspecified role neutral component neutral component 14 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7G IgM-BCR gains access to GM1 gangliosides (CTB) only upon B cell activation. specific antigen-NIP, pervanadate (Perv) and Lat-A - activators of the BCR. ctxb-ighm-1 2 22 unspecified role unspecified role neutral component neutral component 14 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 7G-2 On resting B cells, GM1 gangliosides (CTB) are found in close proximity to the IgD-BCR; IgM-BCR gains access to GM1 gangliosides (CTB) only upon B cell activation. specific antigen-NIP, pervanadate (Perv) and Lat-A - activators of the BCR. ighm-ighm-3 10 19 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 5G, Figure 7A Supplementary 1 A clear difference was shown between the relative IgM:IgM conformation or distance between resting (positive result) and activated B cells (negative result). The Fab-PLA signal is lost upon exposure of B cells to antigen (NIP), or in presence of BCR-activating drugs. Specific antigen- NIP, pervanadate and Lat-A. More NIP is required to lose the positive PLA signal after Syk inhibitor R406 treatment. ighm-ighm-4 8 22 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7D Supplementary 1 The Fab-PLA signal is lost upon exposure of B cells to antigen (NIP), or in presence of BCR-activating drugs. Specific antigen- NIP, pervanadate and Lat-A - activators of the BCR ighdm-ighdm-4 8 12 unspecified role unspecified role neutral component neutral component 22 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7E Supplementary 1 Resting TKO-D cells display a close proximity between the IgD-BCR and the CD19/CD20 module but lose this interaction upon BCR stimulation The Fab-PLA signal is lost upon exposure of B cells to antigen (NIP), or in presence of BCR-activating drugs. Specific antigen- NIP, pervanadate and Lat-A - activators of the BCR cd19-ighdm-2 8 22 unspecified role unspecified role neutral component neutral component 20 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7F Supplementary 1 Resting TKO-D cells display a close proximity between the IgD-BCR and the CD19/CD20 module but lose this interaction upon BCR stimulation The Fab-PLA signal is lost upon exposure of B cells to antigen (NIP), or in presence of BCR-activating drugs. Specific antigen- NIP, pervanadate and Lat-A - activators of the BCR ighm-ighm-5 10 19 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7B Supplementary 1 IgM-BCR only gains contact to the CD19/CD20 module upon B cell activation. Specific antigen- NIP, pervanadate and Lat-A- - activators of the BCR cd19-ighm-7 10 20 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7C Supplementary 1 IgM-BCR only gains contact to the CD19/CD20 module upon B cell activation. Specific antigen- NIP, pervanadate and Lat-A- - activators of the BCR cd19-ighm-8 10 13 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7B Supplementary 2 Specific antigen- NIP. ighdm-ighdm-1 8 22 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 7 Supplementary 2 There is slight loss of Fab-PLA signal upon exposure of B cells to antigen (NIP). Specific antigen- NIP ighm-ighm-11 3 19 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line physical association physical association 2B