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/ Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Immunity 2014 imex curation Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu 21-10-2015: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-t-cell Mouse T cell t-cell Peripheral blood T-lymphocytes ch-ip chromatin immunoprecipitation assay ch-ip q-pcr quantitative pcr q-pcr Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D Naive, Th0 (precursor) or Induced TReg cells:(Naïve -CD44loCD62L+CD25– CD4+ T cells). Cells were stimulated with plate-bound anti-CD3, anti-CD28 and appropriate cytokines. The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu Accepted 2015-SEP-11 by ORCHARD Immunity 2014 Only protein-protein interactions imex curation imex curation Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Immunity 2014 imex curation Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu 21-10-2015: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-t-cell Mouse T cell t-cell Peripheral blood T-lymphocytes anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D WT, and Cd44–/– or or Lgals9–/– iTreg cells. The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu Accepted 2015-SEP-11 by ORCHARD Immunity 2014 Only protein-protein interactions imex curation imex curation Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Immunity 2014 imex curation Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu 21-10-2015: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-t-cell Mouse T cell t-cell Peripheral blood T-lymphocytes pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D iTreg cells from WT or Lgals9–/– mice. The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu Accepted 2015-SEP-11 by ORCHARD Immunity 2014 Only protein-protein interactions imex curation imex curation Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Immunity 2014 imex curation Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu 21-10-2015: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-t-cell Mouse T cell t-cell Peripheral blood T-lymphocytes confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Wu C., Thalhamer T., Franca RF., Xiao S., Wang C., Hotta C., Zhu C., Hirashima M., Anderson AC., Kuchroo VK. IBD - Inflammatory bowel disease causality from Figs 1C, S1C, S2D/E, 2F/G, 6D,7B, Supp 7D ....supp6D Th0 (precursor) and iTreg cells from WT or Lgals9–/– mice. Cells were stimulated with anti-CD3 and anti-CD28, in the presence or absence of TGF-beta. The authors have stated that TGF-beta is of human origin. vkuchroo@rics.bwh.harvard.edu Accepted 2015-SEP-11 by ORCHARD Immunity 2014 Only protein-protein interactions imex curation imex curation leg9_mouse_gene Galectin-9 Lgals9 gene gene mouse Mus musculus Mouse tgfr1_mouse TGF-beta receptor type-1 Tgfbr1 Transforming growth factor-beta receptor type I ESK2 protein protein mouse Mus musculus Mouse MEAAAAAPRRPQLLIVLVAAATLLPGAKALQCFCHLCTKDNFTCETDGLCFVSVTETTDKVIHNSMCIAEIDLIPRDRPFVCAPSSKTGAVTTTYCCNQDHCNKIELPTTGPFSEKQSAGLGPVELAAVIAGPVCFVCIALMLMVYICHNRTVIHHRVPNEEDPSLDRPFISEGTTLKDLIYDMTTSGSGSGLPLLVQRTIARTIVLQESIGKGRFGEVWRGKWRGEEVAVKIFSSREERSWFREAEIYQTVMLRHENILGFIAADNKDNGTWTQLWLVSDYHEHGSLFDYLNRYTVTVEGMIKLALSTASGLAHLHMEIVGTQGKPAIAHRDLKSKNILVKKNGTCCIADLGLAVRHDSATDTIDIAPNHRVGTKRYMAPEVLDDSINMKHFESFKRADIYAMGLVFWEIARRCSIGGIHEDYQLPYYDLVPSDPSVEEMRKVVCEQKLRPNIPNRWQSCEALRVMAKIMRECWYANGAARLTALRIKKTLSQLSQQEGIKM DIP protein Q64729 original sequence version: 120 DIP protein Q64729 original sequence version: 131 DIP protein Q64729 original sequence version: 157 BB8BB6D2261793AF foxo3_mouse_gene Forkhead box protein O3 Foxo3 Fkhr2 Foxo3a gene gene mouse Mus musculus Mouse h4_mouse Histone H4 H4c11 H4c12 H4c14 H4c2 H4c3 H4c4 H4c6 H4c8 H4c9 H4f16 Hist1h4a Hist4h4 Hist1h4b Hist1h4c Hist1h4d Hist1h4f Hist1h4h Hist1h4i Hist1h4j Hist1h4k Hist2h4a Hist2h4 Hist1h4m H4-53 H4-12 H4c1 protein protein mouse Mus musculus Mouse MSGRGKGGKGLGKGGAKRHRKVLRDNIQGITKPAIRRLARRGGVKRISGLIYEETRGVLKVFLENVIRDAVTYTEHAKRKTVTAMDVVYALKRQGRTLYGFGG DIP protein P62806 original sequence version: 108 A9E5DFD3F8B97598 leg9_mouse Galectin-9 Lgals9 protein protein mouse Mus musculus Mouse MALFSAQSPYINPIIPFTGPIQGGLQEGLQVTLQGTTKSFAQRFVVNFQNSFNGNDIAFHFNPRFEEGGYVVCNTKQNGQWGPEERKMQMPFQKGMPFELCFLVQRSEFKVMVNKKFFVQYQHRVPYHLVDTIAVSGCLKLSFITFQNSAAPVQHVFSTLQFSQPVQFPRTPKGRKQKTQNFRPAHQAPMAQTTIHMVHSTPGQMFSTPGIPPVVYPTPAYTIPFYTPIPNGLYPSKSIMISGNVLPDATRFHINLRCGGDIAFHLNPRFNENAVVRNTQINNSWGQEERSLLGRMPFSRGQSFSVWIICEGHCFKVAVNGQHMCEYYHRLKNLQDINTLEVAGDIQLTHVQT Binds galactosides. May play a role in thymocyte-epithelial interactions relevant to the biology of the thymus. Is a ligand for HAVCR2/TIM3. Induces T-helper type 1 lymphocyte (Th1) death mint B54036F6E280C531 cd44_mouse CD44 antigen Phagocytic glycoprotein I Cd44 Ly-24 Phagocytic glycoprotein 1 HUTCH-I Extracellular matrix receptor III GP90 lymphocyte homing/adhesion receptor Hermes antigen Hyaluronate receptor Lymphocyte antigen 24 protein protein mouse Mus musculus Mouse MDKFWWHTAWGLCLLQLSLAHQQIDLNVTCRYAGVFHVEKNGRYSISRTEAADLCQAFNSTLPTMDQMKLALSKGFETCRYGFIEGNVVIPRIHPNAICAANHTGVYILVTSNTSHYDTYCFNASAPPEEDCTSVTDLPNSFDGPVTITIVNRDGTRYSKKGEYRTHQEDIDASNIIDDDVSSGSTIEKSTPEGYILHTYLPTEQPTGDQDDSFFIRSTLATIASTVHSKSHAAAQKQNNWIWSWFGNSQSTTQTQEPTTSATTALMTTPETPPKRQEAQNWFSWLFQPSESKSHLHTTTKMPGTESNTNPTGWEPNEENEDETDTYPSFSGSGIDDDEDFISSTIATTPRVSARTEDNQDWTQWKPNHSNPEVLLQTTTRMADIDRISTSAHGENWTPEPQPPFNNHEYQDEEETPHATSTTPNSTAEAAATQQETWFQNGWQGKNPPTPSEDSHVTEGTTASAHNNHPSQRITTQSQEDVSWTDFFDPISHPMGQGHQTESKDTDSSHSTTLQPTAAPNTHLVEDLNRTGPLSVTTPQSHSQNFSTLHGEPEEDENYPTTSILPSSTKSSAKDARRGGSLPTDTTTSVEGYTFQYPDTMENGTLFPVTPAKTEVFGETEVTLATDSNVNVDGSLPGDRDSSKDSRGSSRTVTHGSELAGHSSANQDSGVTTTSGPMRRPQIPEWLIILASLLALALILAVCIAVNSRRRCGQKKKLVINGGNGTVEDRKPSELNGEASKSQEMVHLVNKEPSETPDQCMTADETRNLQSVDMKIGV Main cell surface receptor for hyaluronate. Adhesion to mucosal high endothelial venule and to types I and VI collagen. Probably involved in matrix adhesion, lymphocyte activation and lymph node homing mint BD2C073250F6C956 smad3_mouse Mothers against decapentaplegic homolog 3 Smad3 Madh3 SMAD family member 3 protein protein mouse Mus musculus Mouse MSSILPFTPPIVKRLLGWKKGEQNGQEEKWCEKAVKSLVKKLKKTGQLDELEKAITTQNVNTKCITIPRSLDGRLQVSHRKGLPHVIYCRLWRWPDLHSHHELRAMELCEFAFNMKKDEVCVNPYHYQRVETPVLPPVLVPRHTEIPAEFPPLDDYSHSIPENTNFPAGIEPQSNIPETPPPGYLSEDGETSDHQMNHSMDAGSPNLSPNPMSPAHNNLDLQPVTYCEPAFWCSISYYELNQRVGETFHASQPSMTVDGFTDPSNSERFCLGLLSNVNRNAAVELTRRHIGRGVRLYYIGGEVFAECLSDSAIFVQSPNCNQRYGWHPATVCKIPPGCNLKIFNNQEFAALLAQSVNQGFEAVYQLTRMCTIRMSFVKGWGAEYRRQTVTSTPCWIELHLNGPLQWLDKVLTQMGSPSIRCSSVS DIP protein Q8BUN5 original sequence version: 57 46DF5E8B371321AC smad3-lgals9-1 1 5 unspecified role unspecified role prey prey multiple_regions sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait association association 2C Q8BUN5 increases expression at RNA level of ENSMUSG00000001123 Q8BUN5 increases expression at protein level of ENSMUSG00000001123 This interaction was not seen in Th0 (precursor) cells. P01137 increases expression at protein level of ENSMUSG00000001123 P01137 increases expression at RNA level of ENSMUSG00000001123 the tgf-beta1 caus. statements have been taken from Supp figs 2B/C - main text states TGF-beta was used, even if the figure text says nowt. smad3-lgals9-2 4 10 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization 5A This interaction was only seen in cells from WT mice, and not from cells of Lgals9–/– mice. TGF-beta it is not stated for Fig 5A that co-loc is at the "plasma-membrane" - however it IS for Fig 5C - which looks to occur at the same location as here;(T cells do not have much cytoplasm so the DAPI staining is not relevant!!) cd44-tgfbr1-1 2 10 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association 5A This interaction was only seen in cells from WT mice, and not from cells of Lgals9–/– mice. cd44-tgfbr1-2 2 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association 5A-2 This interaction was only seen in cells from WT mice, and not from cells of Lgals9–/– mice. cd44-tgfbr1-3 3 10 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 5C foxo3-smad3-1 1 7 unspecified role unspecified role prey prey cns1_enhancer__region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait association association 6A, Supp 6B, (S1C) O00182-2 increases expression at protein level of ENSMUSG00000048756 Q8BUN5 increases expression at RNA level of ENSMUSG00000048756 Q8BUN5 increases expression at protein level of ENSMUSG00000048756 O08573 increases binding of Q8BUN5 to ENSMUSG00000048756 P15379 is required for the expression at protein level of ENSMUSG00000048756 A substantial synergistic effect of galectin-9 (O00182-2) with TGF-beta in inducing Foxp3 expression was observed. Lactose inhibited this expression. O08573 is required for binding of Q8BUN5 to ENSMUSG00000048756 No binding of Smad3 to Foxp3 CNS1 region is seen in Th0 (precursor) cells. O00182-2 increases expression at RNA level of ENSMUSG00000048756 P01137 increases expression at protein level of ENSMUSG00000048756 CHEBI:17716 inhibits P01137 and O00182-2 induced increase in expression at protein level of ENSMUSG00000048756 CHEBI:17716 indirectly decreases expression at protein level of ENSMUSG00000048756 I have added two causality statements for the effect of lactose on foxp3 expression (FACS) - "CHEBI:17716 inhibits P01137 and O00182-2 induced increase in expression at protein level of ENSMUSG00000048756" - is not on the list of "previously used" statements - so please deleted if you dont think it is OK.. I have also added an "indirect" effect statement hist1h4h-foxo3-1 1 7 unspecified role unspecified role prey prey cns1_enhancer__region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait region ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 6 A/B O08573 increases binding of P62806 to ENSMUSG00000048756 O00182-2 increases binding of P62806 to ENSMUSG00000048756 TGF-beta and galectin-9 P01137 increases binding of P62806 to ENSMUSG00000048756 cd44-tgfbr1-4 2 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Supp 5C cd44-lgals9-1 2 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association Supp 5C-2