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/ Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Cancer Cell (1535-6108) 2010 imex curation Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions human-22rv1 Homo sapiens 22rv1 Human prostate adenocarcinoma cell line Also known as CWR22Rv1 or Rv1. Cells dsplay a NE (neuroendocrine phenotype). ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Cells were maintained in 1% Oxygen for 5 or 6 hrs. Cells were crosslinked using 1% formaldehyde for 10 min at RT. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 jyoti Only protein-protein interactions imex curation Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Cancer Cell (1535-6108) 2010 imex curation Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions mouse-tramp_c Mouse TRAMP-C tramp_c Mouse TRAMP-C cell line Tissue was obtained from a 32-week-old prostatic adenocarcinoma from a C57BL16male TRAMP mouse. (TRansgenic Adenocarcinoma Mouse Prostate.) TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter driving prostate-specific epithelial expression of the SV40 large T antigen. Tissue was obtained from a 32-week-old prostatic adenocarcinoma from a C57BL16male TRAMP mouse. (TRansgenic Adenocarcinoma Mouse Prostate.) TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter driving prostate-specific epithelial expression of the SV40 large T antigen. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Cells were maintained in 1% Oxygen for 5 or 6 hrs. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 jyoti Only protein-protein interactions imex curation Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Cancer Cell (1535-6108) 2010 imex curation Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions mouse-tramp_c Mouse TRAMP-C tramp_c Mouse TRAMP-C cell line Tissue was obtained from a 32-week-old prostatic adenocarcinoma from a C57BL16male TRAMP mouse. (TRansgenic Adenocarcinoma Mouse Prostate.) TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter driving prostate-specific epithelial expression of the SV40 large T antigen. Tissue was obtained from a 32-week-old prostatic adenocarcinoma from a C57BL16male TRAMP mouse. (TRansgenic Adenocarcinoma Mouse Prostate.) TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter driving prostate-specific epithelial expression of the SV40 large T antigen. ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Cells were maintained in 1% Oxygen for 5 or 6 hrs. Cells were crosslinked using 1% formaldehyde for 10 min at RT. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 jyoti Only protein-protein interactions imex curation Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Cancer Cell (1535-6108) 2010 imex curation Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 jyoti Only protein-protein interactions imex curation Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. Cancer Cell (1535-6108) 2010 imex curation Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Qi J., Nakayama K., Cardiff RD., Borowsky AD., Kaul K., Williams R., Krajewski S., Mercola D., Carpenter PM., Bowtell D., Ronai ZA. Cancer - Interactions investigated in the context of cancer Originally I thought Participant Detection was "anti-tag Western" - but I now cannot find anything to confirm this. Unfortunately there is no mention of an 'anti-FOXA' antibody (as stated to be used on the image) or an antibodies to various other proteins either. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 jyoti Only protein-protein interactions imex curation arnt_mouse Aryl hydrocarbon receptor nuclear translocator Arnt Dioxin receptor, nuclear translocator Hypoxia-inducible factor 1-beta protein protein mouse Mus musculus Mouse MAATTANPEMTSDVPSLGPTIASGNPGPGIQGGGAVVQRAIKRRSGLDFDDEVEVNTKFLRCDDDQMCNDKERFARSDDEQSSADKERLARENHSEIERRRRNKMTAYITELSDMVPTCSALARKPDKLTILRMAVSHMKSLRGTGNTSTDGSYKPSFLTDQELKHLILEAADGFLFIVSCETGRVVYVSDSVTPVLNQPQSEWFGSTLYDQVHPDDVDKLREQLSTSENALTGRVLDLKTGTVKKEGQQSSMRMCMGSRRSFICRMRCGTSSVDPVSMNRLSFLRNRCRNGLGSVKEGEPHFVVVHCTGYIKAWPPAGVSLPDDDPEAGQGSKFCLVAIGRLQVTSSPNCTDMSNICQPTEFISRHNIEGIFTFVDHRCVATVGYQPQELLGKNIVEFCHPEDQQLLRDSFQQVVKLKGQVLSVMFRFRSKTREWLWMRTSSFTFQNPYSDEIEYIICTNTNVKNSSQEPRPTLSNTIPRSQLGPTANLSLEMGTGQLPSRQQQQQHTELDMVPGRDGLASYNHSQVSVQPVASAGSEHSKPLEKSEGLFAQDRDPRFPEIYPSITADQSKGISSSTVPATQQLFSQGSSFPPNPRPAENFRNSGLTPPVTIVQPSSSAGQILAQISRHSNPAQGSAPTWTSSSRPGFAAQQVPTQATAKTRSSQFGVNNFQTSSSFSAMSLPGAPTASSGTAAYPALPNRGSNFPPETGQTTGQFQARTAEGVGVWPQWQGQQPHHRSSSSEQHVQQTQAQAPSQPEVFQEMLSMLGDQSNTYNNEEFPDLTMFPPFSE DIP protein P53762 original sequence version: 78 DIP protein P53762-1 original sequence version: 143 BDEC0991CD57D402 hes6_human_gene Human HES6 gene gene gene human Homo sapiens Human foxa2_mouse Hepatocyte nuclear factor 3-beta Foxa2 Hnf3b Tcf-3b Tcf3b Forkhead box protein A2 protein protein mouse Mus musculus Mouse MLGAVKMEGHEPSDWSSYYAEPEGYSSVSNMNAGLGMNGMNTYMSMSAAAMGGGSGNMSAGSMNMSSYVGAGMSPSLAGMSPGAGAMAGMSGSAGAAGVAGMGPHLSPSLSPLGGQAAGAMGGLAPYANMNSMSPMYGQAGLSRARDPKTYRRSYTHAKPPYSYISLITMAIQQSPNKMLTLSEIYQWIMDLFPFYRQNQQRWQNSIRHSLSFNDCFLKVPRSPDKPGKGSFWTLHPDSGNMFENGCYLRRQKRFKCEKQLALKEAAGAASSGGKKTAPGSQASQAQLGEAAGSASETPAGTESPHSSASPCQEHKRGGLSELKGAPASALSPPEPAPSPGQQQQAAAHLLGPPHHPGLPPEAHLKPEHHYAFNHPFSINNLMSSEQQHHHSHHHHQPHKMDLKAYEQVMHYPGGYGSPMPGSLAMGPVTNKAGLDASPLAADTSYYQGVYSRPIMNSS 2F20F15AB372F557 ep300_human Histone acetyltransferase p300 Histone butyryltransferase p300 Histone crotonyltransferase p300 Protein propionyltransferase p300 EP300 P300 E1A-associated protein p300 Protein 2-hydroxyisobutyryltransferase p300 Protein lactyltransferas p300 protein protein human Homo sapiens Human MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDINQLQTSLGMVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQVMASQAQQSSPGLGLINSMVKSPMTQAGLTSPNMGMGTSGPNQGPTQSTGMMNSPVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRQNMQYPNPGMGSAGNLLTEPLQQGSPQMGGQTGLRGPQPLKMGMMNNPNPYGSPYTQNPGQQIGASGLGLQIQTKTVLSNNLSPFAMDKKAVPGGGMPNMGQQPAPQVQQPGLVTPVAQGMGSGAHTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQPILTGAPVGLGNPSSLGVGQQSAPNLSTVSQIDPSSIERAYAALGLPYQVNQMPTQPQVQAKNQQNQQPGQSPQGMRPMSNMSASPMGVNGGVGVQTPSLLSDSMLHSAINSQNPMMSENASVPSLGPMPTAAQPSTTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAAGMVPVSMNPGPNMGQPQPGMTSNGPLPDPSMIRGSVPNQMMPRITPQSGLNQFGQMSMAQPPIVPRQTPPLQHHGQLAQPGALNPPMGYGPRMQQPSNQGQFLPQTQFPSQGMNVTNIPLAPSSGQAPVSQAQMSSSSCPVNSPIMPPGSQGSHIHCPQLPQPALHQNSPSPVPSRTPTPHHTPPSIGAQQPPATTIPAPVPTPPAMPPGPQSQALHPPPRQTPTPPTTQLPQQVQPSLPAAPSADQPQQQPRSQQSTAASVPTPTAPLLPPQPATPLSQPAVSIEGQVSNPPSTSSTEVNSQAIAEKQPSQEVKMEAKMEVDQPEPADTQPEDISESKVEDCKMESTETEERSTELKTEIKEEEDQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQGESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAGFVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETRWHCTVCEDYDLCITCYNTKNHDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSIQRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPICKQLIALCCYHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQRTGVVGQQQGLPSPTPATPTTPTGQQPTTPQTPQPTSQPQPTPPNSMPPYLPRTQAAGPVSQGKAAGQVTPPTPPQTAQPPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQMPPMTPMAPMGMNPPPMTRGPSGHLEPGMGPTGMQQQPPWSQGGLPQPQQLQSGMPRPAMMSVAQHGQPLNMAPQPGLGQVGISPLKPGTVSQQALQNLLRTLRSPSSPLQQQQVLSILHANPQLLAAFIKQRAAKYANSNPQPIPGQPGMPQGQPGLQPPTMPGQQGVHSNPAMQNMNPMQAGVQRAGLPQQQPQQQLQPPMGGMSPQAQQMNMNHNTMPSQFRDILRRQQMMQQQQQQGAGPGIGPGMANHNQFQQPQGVGYPPQQQQRMQHHMQQMQQGNMGQIGQLPQALGAEAGASLQAYQQRLLQQQMGSPVQPNPMSPQQHMLPNQAQSPHLQGQQIPNSLSNQVRSPQPVPSPRPQSQPPHSSPSPRMQPQPSPHHVSPQTSSPHPGLVAAQANPMEQGHFASPDQNSMLSQLASNPGMANLHGASATDLGLSTDNSDLNSNLSQSTLDIH DIP protein Q09472 original sequence version: 193 DIP protein Q09472 original sequence version: 186 DIP protein Q09472 original sequence version: 107 DIP protein Q09472 original sequence version: 112 DIP protein Q09472 original sequence version: 181 DIP protein Q09472 original sequence version: 183 DIP protein Q09472 original sequence version: 136 DIP protein Q09472 original sequence version: 146 DIP protein Q09472 original sequence version: 165 DIP protein Q09472 original sequence version: 192 DIP protein Q09472 original sequence version: 200 DIP protein Q09472 original sequence version: 206 DIP protein Q09472 original sequence version: 194 DIP protein Q09472 original sequence version: 171 DIP protein Q09472 original sequence version: 142 DIP protein Q09472 original sequence version: 180 8E869E1F174A6FEB q8bj14_mouse Ep300 protein protein mouse Mus musculus Mouse MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDISQLQTSLGIVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQAMASQAQQNSPGLSLINSMVKSPMAQTGLTSPNMGIGSSGPNQGPTQSPAGMMNSPVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRPNMQYPNAGMGNAGSLLTEPLQQGSPQMGGQPGLRGPQPLKMGMMNNPSPYGSPYTQNSGQQIGASGLGLQIQTKTVLPNNLSPFAMDKKAVPGGGMPSMGQQPTPSVQQPGLVTPVAAGMGSGAHTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQSILTGAPVGLGNPSSLGVGQQSTPSLSTVSQIDPSSIERAYAALGLPYQVNQIPPQPQVQAKNQQSQPSGQSPQGMRSVNNMSASPMGVNGGVGVQTPNLLSDSMLHSTINSQNPMMSENAGVASLGPLPTAAQPSSTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAPGMGPVPMNTGSNMGQQPTGMTTNGPVPDPSMIRGSVPNHMMPRMTPQPGNV C382B614A837CE49 hif1a_mouse Hypoxia-inducible factor 1-alpha Hif1a ARNT-interacting protein protein protein mouse Mus musculus Mouse MEGAGGENEKKKMSSERRKEKSRDAARSRRSKESEVFYELAHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGGLDSEDEMKAQMDCFYLKALDGFVMVLTDDGDMVYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGPVRKGKELNTQRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNYVVSGIIQHDLIFSLQQTESVLKPVESSDMKMTQLFTKVESEDTSCLFDKLKKEPDALTLLAPAAGDTIISLDFGSDDTETEDQQLEDVPLYNDVMFPSSNEKLNINLAMSPLPSSETPKPLRSSADPALNQEVALKLESSPESLGLSFTMPQIQDQPASPSDGSTRQSSPERLLQENVNTPNFSQPNSPSEYCFDVDSDMVNVFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESNSPSPPSMSTVTGFQQTQLQKPTITATATTTATTDESKTETKDNKEDIKILIASPSSTQVPQETTTAKASAYSGTHSRTASPDRAGKRVIEQTDKAHPRSLNLSATLNQRNTVPEEELNPKTIASQNAQRKRKMEHDGSLFQAAGIGTLLQQPGDCAPTMSLSWKRVKGFISSEQNGTEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN 84D64ECAC2CC753B kdm3a_mouse_gene Mouse Jmjd1a/2a gene gene gene mouse Mus musculus Mouse hes6_mouse_gene Mouse HES6 gene gene gene mouse Mus musculus Mouse foxa2_rat Hepatocyte nuclear factor 3-beta Foxa2 Hnf3b Tcf-3b Tcf3b Forkhead box protein A2 protein protein rat Rattus norvegicus (Rat) Rat MLGAVKMEGHEPSDWSSYYAEPEGYSSVSNMNASLGMNGMNTYMSMSAAAMGSGSGNMSAGSMNMSSYVGAGMSPSLAGMSPGAGAMAGMSGSAGAAGVAGMGPHLSPSLSPLGGQAAGAMGGLAPYANMNSMSPMYGQAGLSRARDPKTYRRSYTHAKPPYSYISLITMAIQQSPNKMLTLSEIYQWIMDLFPFYRQNQQRWQNSIRHSLSFNDFLKVPRAPDKPGKGSFWTLHPDSGNMFENGCYLRRQKRFKCENELALKEAAGAGSGGGKKTAPGTQASQVQLGEAAGSASETPAGTESPHSSASPCQEHKRGGLSELKGTPASALSPPEPAPSPGQQQQAAAHLLGPPHHPGLPPEAHLKPEHHYAFNHPFSINNLMSSEQQHHHSHHHHQPHKMDLKTYEQVMHYPGGYGSPMPGSLAMGPVTNKAGLDASPLAADTSYYQGVYSRPIMNSS 9FF754142C194055 vegfa_human_gene Human VEGFA gene gene gene human Homo sapiens Human kdm3a_human_gene Human Jmjd1a/2a gene gene gene human Homo sapiens Human vegfa_mouse_gene Mouse VEGFA gene gene gene mouse Mus musculus Mouse hif1a-foxa2 4 14 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association HA-FoxA2, but not HA-FoxA1, was identified as an HIF-associated protein. Fig 3E. Species:HA-FOXA2 is only ever mentioned in Supp Exp Methods as Rat and Flag-HIF a1 as Mouse. hif1a-foxa2-arnt 2 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey association association Endogenous FoxA2 was coprecipitated with endogenous HIF-1a (Figure 3F) or HIF-1b (Figure 3G) in TRAMP-C cells maintained under hypoxia. Fig 3F. arnt-hif1a-foxa2 2 6 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey association association Endogenous FoxA2 was coprecipitated with endogenous HIF-1a (Figure 3F) or HIF-1b (Figure 3G) in TRAMP-C cells maintained under hypoxia. Fig 3G. The association of HIF-1b with FoxA2 was abolished following HIF-1a knockdown (Figure 3G). his-foxa2-hif1a 5 8 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 11 unspecified role unspecified role prey prey bhlh-pas domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association In vitro binding using truncation mutants of HIF-a (Figure S2C) and FoxA2 (Figure S2B) identified the bHLH-PAS domain of HIF-1a (Figure 3H) and the N-TAD of FoxA2 (Figure 3I) as the minimal regions mediating the HIF-1a-FoxA2 interaction. Fig 3H. Domains of HIF1a - as stated in Fig S2C, do not correspond to the Interpro domain a.a. numbers well enough to enter any XRef. No "expressed in" for His -FoxA2. Species assummed "Murine" for "Flag-HIF-1a truncation mutants" - See Supplement:(Mouse HIF-1a and HIF-1a 531-822 were subcloned into T vector for in vitro translation of proteins without Tag. Flag-HIF-1a truncation mutants in T vector were obtained from Dr. Lorenz Poellinger (Karolinska Institute, Stockholm, Sweden) and used for in vitro protein translation.) hifa1-foxa2 5 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-tad domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 11 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association In vitro binding using truncation mutants of HIF-1a (Figure S2C) and FoxA2 (Figure S2B) identified the bHLH-PAS domain of HIF-1a (Figure 3H) and the N-TAD of FoxA2 (Figure 3I) as the minimal regions mediating the HIF-1a-FoxA2 interaction. Fig 3I. N_TAD domain amino acid numbers do not seem to be stated anywhere in the paper. Species of FoxA2 truncation mutants is not explicitly stated, but I assume it is Murine. (subcloned into pcDNA3 vector with an N-terminal myc Tag - murine) InterPro gives N_TAD domain as aas 17-158. Authors state the M1 construct is aas 1-185 and M2 as 1-265. So I have entered the M1 construct as "sufficient to bind" and left the Interpro XRef in place, though it is a little shorter than the construct used. hif1a-foxa2-arnt-1 2 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey association association Although the bHLH-PAS domain of HIF-1a was found to interact with FoxA2 and the PAS domain of HIF-1a is known to interact with HIF-1b, alternation of the FoxA2 level in vitro or in TRAMP-C cells had no apparent effect on the interaction of HIF-1b with HIF-1a (Figures 3J and 3K). Fig 3J. hif1a-foxa2-hif1b 5 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 11 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Fig 3K. I do not think the HIF1-b has a tag. (but may have missed one...) Although the bHLH-PAS domain of HIF-1a was found to interact with FoxA2 and the PAS domain of HIF-1a is known to interact with HIF-1b alternation of the FoxA2 level in vitro or in TRAMP-C cells had no apparent effect on the interaction of HIF-1b with HIF-1a. I think that the "Increasing levels of FoxA2" i.e. "alternation of the FoxA2 level" are shown by an solid horizontal black triangle on the image, at the top - no numerical values are given for FoxA2 levels. - I have not added any comment/annotation to show this, mainly as I cannot think of any sensible way of stating this! hif1a-hes6 3 13 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait association association Fig 4E, Fig S3B (lanes 3). ChIP assays of the HRE-containing region of Hes6 promoter were carried out. Binding was observed to the HRE located at 66 bp upstream of Hes6 transcriptional start site. foxa2-hes6 3 8 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey association association Fig 4E, Fig S3B (lanes 5). ChIP assays of the HRE-containing region of Hes6 promoter were carried out. Binding was observed to the HRE located at 66 bp upstream of Hes6 transcriptional start site. hif1b-hes6 3 6 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey association association Fig S3B (lanes 4). ChIP assays of the HRE-containing region of Hes6 promoter were carried out. Binding was observed to the HRE located at 66 bp upstream of Hes6 transcriptional start site. p300-hes6-1 3 10 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey association association Fig 4E. ChIP assays of the HRE-containing region of Hes6 promoter were carried out. Binding was observed to the HRE located at 66 bp upstream of Hes6 transcriptional start site. Binding of p300 to the Hes6 promoter was attenuated after FoxA2 knockdown. I had some difficulty finding an appropriate entry for "p300" - I settled on this one as the human version states it interacts with HIF1a. Q8BJ14 or B2RWS6 (or Q3USJ1? ) are all possible entries for the same Gene ID - I ruled out the last one as it is very short - but p300 could be to either of the other Trembl entries. B2RWS6 is not mentioned in Entrez gene (no Ref-Seq entry)- so I entered the other Trembl entry as Bait. -there is a X3-X4 difference in length and searching for ENSMUSG00000055024 does not give any SwissP result. p300-hes6-2 3 13 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait association association Fig 4F. FoxA2 knockdown also reduced the binding of p300 to HRE-containing promoters of Jmjd1a and VEGFA (and Hes6) in both TRAMP-C cells and Rv1 cells. I had some difficulty finding an appropriate entry for "p300" - I settled on this one as the human version states it interacts with HIF1a. Q8BJ14 or B2RWS6 (or Q3USJ1? ) are all possible entries for the same Gene ID - I ruled out the last one as it is very short - but p300 could be to either of the other Trembl entries. B2RWS6 is not mentioned in Entrez gene (no Ref-Seq entry)- so I entered the other Trembl entry as Bait. -there is a X3-X4 difference in length and searching for ENSMUSG00000055024 does not give any SwissP result. p300-vegfa 3 17 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait association association Fig 4F. FoxA2 knockdown also reduced the binding of p300 to HRE-containing promoters of Jmjd1a and VEGFA (and Hes6) in both TRAMP-C cells and Rv1 cells. I had some difficulty finding an appropriate entry for "p300" - I settled on this one as the human version states it interacts with HIF1a. Q8BJ14 or B2RWS6 (or Q3USJ1? ) are all possible entries for the same Gene ID - I ruled out the last one as it is very short - but p300 could be to either of the other Trembl entries. B2RWS6 is not mentioned in Entrez gene (no Ref-Seq entry)- so I entered the other Trembl entry as Bait. -there is a X3-X4 difference in length and searching for ENSMUSG00000055024 does not give any SwissP result. p300-kdma 3 10 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey association association Fig 4F. FoxA2 knockdown also reduced the binding of p300 to HRE-containing promoters of Jmjd1a and VEGFA (and Hes6) in both TRAMP-C cells and Rv1 cells. I had some difficulty finding an appropriate entry for "p300" - I settled on this one as the human version states it interacts with HIF1a. Q8BJ14 or B2RWS6 (or Q3USJ1? ) are all possible entries for the same Gene ID - I ruled out the last one as it is very short - but p300 could be to either of the other Trembl entries. B2RWS6 is not mentioned in Entrez gene (no Ref-Seq entry)- so I entered the other Trembl entry as Bait. -there is a X3-X4 difference in length and searching for ENSMUSG00000055024 does not give any SwissP result. p300-vegfa-1 1 15 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait association association Fig 4F. FoxA2 knockdown also reduced the binding of p300 to HRE-containing promoters of Jmjd1a and VEGFA (and Hes6) in both TRAMP-C cells and Rv1 cells. p300-hes6-3 1 9 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association Fig 4F. FoxA2 knockdown also reduced the binding of p300 to HRE-containing promoters of Jmjd1a and VEGFA (and Hes6) in both TRAMP-C cells and Rv1 cells. p300-kdma-1 1 9 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey association association Fig 4F. FoxA2 knockdown also reduced the binding of p300 to HRE-containing promoters of Jmjd1a and VEGFA (and Hes6) in both TRAMP-C cells and Rv1 cells. p300-hif1a-foxa2 5 9 unspecified role unspecified role bait bait d_ch1 p300 Delta_CH1 required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain in vitro In vitro 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain in vitro In vitro association association FoxA2 enhanced HIF1a/p300 interaction but had no effect on p300 binding to the HIF1a 531-822 mutant that cannot associate with FoxA2. When Flag-p300Delta_CH1 (a p300 mutant lacking the CH1 domain, that cannot interact with HIF1a)was used to replace p300, no binding with HIF/FoxA2 complex was observed. Fig 4K, 4L. NO tags for HIF1A I think - "Mouse HIF-1a and HIF1a 531-822 were subcloned into T vector for in vitro translation of proteins without Tag." (from Supp Mat). Fox A2 DOES have a tag- confirmed by authors. I have entered the HIF1A aas 1-530 feature as "required to bind", as deletion analysis shows that it does not bind if not present. - though i am not sure if this is correct usage of the term "deletion analysis" and the sequence (1-530) is rather larger than i would have expected for "deletion analysis". p300-hif531-822 5 9 unspecified role unspecified role bait bait in vitro In vitro 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association FoxA2 enhanced HIF1a/p300 interaction but had no effect on p300 binding to the HIF1a 531-822 mutant that cannot associate with FoxA2. Fig 4K. NO tags for HIF1A I think - "Mouse HIF-1a and HIF1a 531-822 were subcloned into T vector for in vitro translation of proteins without Tag." (from Supp Mat). Fox A2 does have a tag- confirmed by authors.