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/ http://www.ebi.ac.uk/ kl_sener-2014-6 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-3 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-7 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. "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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-8 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-10 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-5 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-1 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. "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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-2 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-4 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-9 B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. 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. "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 Elife 2014 Only protein-protein interactions imex curation imex curation kl_sener-2014-11 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. 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 Elife 2014 Only protein-protein interactions imex curation imex curation ighm_mouse Immunoglobulin heavy constant mu Ighm Igh-6 protein protein mouse Mus musculus Mouse SQSFPNVFPLVSCESPLSDKNLVAMGCLARDFLPSTISFTWNYQNNTEVIQGIRTFPTLRTGGKYLATSQVLLSPKSILEGSDEYLVCKIHYGGKNRDLHVPIPAVAEMNPNVNVFVPPRDGFSGPAPRKSKLICEATNFTPKPITVSWLKDGKLVESGFTTDPVTIENKGSTPQTYKVISTLTISEIDWLNLNVYTCRVDHRGLTFLKNVSSTCAASPSTDILTFTIPPSFADIFLSKSANLTCLVSNLATYETLNISWASQSGEPLETKIKIMESHPNGTFSAKGVASVCVEDWNNRKEFVCTVTHRDLPSPQKKFISKPNEVHKHPPAVYLLPPAREQLNLRESATVTCLVKGFSPADISVQWLQRGQLLPQEKYVTSAPMPEPGAPGFYFTHSILTVTEEEWNSGETYTCVVGHEALPHLVTERTVDKSTGKPTLYNVSLIMSDTGGTCY ksyk_mouse Tyrosine-protein kinase SYK Syk Spleen tyrosine kinase Sykb ptk72 protein protein mouse Mus musculus Mouse MAGSAVDSANHLTYFFGNITREEAEDYLVQGGMTDGLYLLRQSRNYLGGFALSVAHNRKAHHYTIERELNGTYAISGGRAHASPADLCHYHSQEPDGLICLLKKPFNRPPGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGNISRDESEQTVLIGSKTNGKFLIRARDNSGSYALCLLHEGKVLHYRIDRDKTGKLSIPEGKKFDTLWQLVEHYSYKPDGLLRVLTVPCQKIGAQMGHPGSPNAHPVTWSPGGIISRIKSYSFPKPGHKKPAPPQGSRPESTVSFNPYEPTGGPWGPDRGLQREALPMDTEVYESPYADPEEIRPKEVYLDRSLLTLEDNELGSGNFGTVKKGYYQMKKVVKTVAVKILKNEANDPALKDELLAEANVMQQLDNPYIVRMIGICEAESWMLVMEMAELGPLNKYLQQNRHIKDKNIIELVHQVSMGMKYLEESNFVHRDLAARNVLLVTQHYAKISDFGLSKALRADENYYKAQTHGKWPVKWYAPECINYYKFSSKSDVWSFGVLMWEAFSYGQKPYRGMKGSEVTAMLEKGERMGCPAGCPREMYDLMNLCWTYDVENRPGFTAVELRLRNYYYDVVN 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 cd19_mouse B-lymphocyte antigen CD19 Differentiation antigen CD19 Cd19 protein protein mouse Mus musculus Mouse MPSPLPVSFLLFLTLVGGRPQKSLLVEVEEGGNVVLPCLPDSSPVSSEKLAWYRGNQSTPFLELSPGSPGLGLHVGSLGILLVIVNVSDHMGGFYLCQKRPPFKDIWQPAWTVNVEDSGEMFRWNASDVRDLDCDLRNRSSGSHRSTSGSQLYVWAKDHPKVWGTKPVCAPRGSSLNQSLINQDLTVAPGSTLWLSCGVPPVPVAKGSISWTHVHPRRPNVSLLSLSLGGEHPVREMWVWGSLLLLPQATALDEGTYYCLRGNLTIERHVKVIARSAVWLWLLRTGGWIVPVVTLVYVIFCMVSLVAFLYCQRAFILRRKRKRMTDPARRFFKVTPPSGNGTQNQYGNVLSLPTSTSGQAHAQRWAAGLGSVPGSYGNPRIQVQDTGAQSHETGLEEEGEAYEEPDSEEGSEFYENDSNLGQDQVSQDGSGYENPEDEPMGPEEEDSFSNAESYENADEELAQPVGRMMDFLSPHGSAWDPSREASSLGSQSYEDMRGILYAAPQLHSIQSGPSHEEDADSYENMDKSDDLEPAWEGEGHMGTWGTT 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 ighdm_mouse Ig delta chain C region membrane-bound form protein protein mouse Mus musculus Mouse DKKEPDMFLLSECKAPEENEKINLGCLVIGSQPLKISWEPKKSSIVEHVFPSEMRNGNYTMVLQVTVLASELNLNHTCTINKPKRKEKPFKFPESWDSQSSKRVTPTLQAKNHSTEATKAITTKKDIEGAMAPSNLTVNILTTSTHPEMSSWLLCEVSGFFPENIHLMWLGVHSKMKSTNFVTANPTAQPGGTFQTWSVLRLPVALSSSLDTYTCVVEHEASKTKLNASKSLAISGIVNTIQHSCIMDEQSDSYMDLEEENGLWPTMCTFVALFLLTLLYSGFVTFIKVK 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 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 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 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 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 cd52_mouse CAMPATH-1 antigen Cd52 Cdw52 Mb7 Lymphocyte differentiation antigen B7 protein protein mouse Mus musculus Mouse MKSFLLFLTIILLVVIQIQTGSLGQATTAASGTNKNSTSTKKTPLKSGASSIIDAGACSFLFFANTLMCLFYLS ighm-ighm-9 1779250 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. cd79a-ighm-1 1779250 n/a 1779262 unspecified role unspecified role neutral component neutral component n/a 1779263 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. cd19-ighm-5 1779250 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779264 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 7E CD19 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP ighm-ms4a1-1 1779250 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779265 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779250 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779264 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779250 n/a 1779265 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779250 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779267 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779250 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779267 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. ighdm-ighdm-2 1779250 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. ighm-ighm-2 1779251 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. cd19-ighm-1 1779251 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779264 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 7A CD19 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP cd20-ighm-2 1779251 n/a 1779265 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 7A-2 CD20 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP cd19-ighm-4 1779251 n/a 1779264 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779251 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779265 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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-ighm-1 1779251 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. ighm-ms4a1-2 1779252 n/a 1779268 unspecified role unspecified role neutral component neutral component n/a 1779269 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779252 n/a 1779270 unspecified role unspecified role neutral component neutral component n/a 1779268 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 7C CD19 is only found in close association with the activated but not with the resting IgM-BCR. Specific antigen- NIP ighdm-ms4a1-2 1779252 n/a 1779271 unspecified role unspecified role neutral component neutral component n/a 1779269 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779252 n/a 1779270 unspecified role unspecified role neutral component neutral component n/a 1779271 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 7D CD19 and 20 are found together with the resting IgD-BCR and dissociate from this receptor upon BCR activation Specific antigen- NIP ighm-ighm-10 1779252 n/a 1779268 unspecified role unspecified role neutral component neutral component n/a 1779268 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 4D Pervanadate, Lat A;BCR-activating drugs ighd-ighd-1 1779252 n/a 1779271 unspecified role unspecified role neutral component neutral component n/a 1779271 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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. ighm-ighm-3 1779253 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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-5 1779253 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779264 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779253 n/a 1779265 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779253 n/a 1779272 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 7B Supplementary 2 Specific antigen- NIP. ighm-ighm-4 1779254 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779254 n/a 1779264 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 1779254 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779265 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 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 ighdm-ighdm-1 1779254 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779272 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 7 Supplementary 2 There is slight loss of Fab-PLA signal upon exposure of B cells to antigen (NIP). Specific antigen- NIP ighm-1 1779255 n/a 1779261 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line N/A 1779261 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 false false false 2C Two IgM monomers interact. ighm-ighm-1 1779255 N/A 1779261 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line Stoichiometry: 5.0 n/a 1779261 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line Stoichiometry: 5.0 physical association physical association false false false 2C-2 1-PLA detects both soluble forms of IgM equally well - monomers and pentamers. ighm-ighm-6 1779256 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1E Results did not distinguish between resting and antigen (NIP) stimulated cells. ighm-ighm-7 1779257 n/a 1779261 unspecified role unspecified role neutral component neutral component n/a 1779261 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 proximity proximity close-contact colocalization neighbourhood interaction false false false 1D Results did not distinguish between resting and antigen (NIP) stimulated cells. ighm-ighm-8 1779258 n/a 1779261 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line Stoichiometry: 5.0 physical association physical association false false false 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. ighdm-ighdm-3 1779259 n/a 1779266 unspecified role unspecified role neutral component neutral component n/a 1779266 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 proximity proximity close-contact colocalization neighbourhood interaction false false false 3B No oligomerisation of IgD was seen when an IgD mutant was transfected. (exact mutations are not stated.) ighm-ighm-11 1779260 N/A 1779261 unspecified role unspecified role neutral component neutral component mouse-b1-8 Mouse B1-8 b1-8 B1-8 anti-NIP specific hybridoma cell line Stoichiometry: 5.0 physical association physical association false false false 2B