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/ qi-2010a-4 Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. 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 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 Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation qi-2010a-3 Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. 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 Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation qi-2010a-2 Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. 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 Cells were maintained in 1% Oxygen for 5 or 6 hrs. Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation qi-2010a-5 Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. 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 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 Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation qi-2010a-1 Siah2-dependent concerted activity of HIF and FoxA2 regulates formation of neuroendocrine phenotype and neuroendocrine prostate tumors. 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 Species of constructs and proteins supplied by the authors. ronai@sanfordburnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation hes6_mouse_gene Mouse HES6 gene gene gene mouse Mus musculus Mouse hif1a_mouse Hypoxia-inducible factor 1-alpha Hif1a ARNT-interacting protein protein protein mouse Mus musculus Mouse MEGAGGENEKKKMSSERRKEKSRDAARSRRSKESEVFYELAHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGGLDSEDEMKAQMDCFYLKALDGFVMVLTDDGDMVYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGPVRKGKELNTQRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNYVVSGIIQHDLIFSLQQTESVLKPVESSDMKMTQLFTKVESEDTSCLFDKLKKEPDALTLLAPAAGDTIISLDFGSDDTETEDQQLEDVPLYNDVMFPSSNEKLNINLAMSPLPSSETPKPLRSSADPALNQEVALKLESSPESLGLSFTMPQIQDQPASPSDGSTRQSSPERLLQENVNTPNFSQPNSPSEYCFDVDSDMVNVFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESNSPSPPSMSTVTGFQQTQLQKPTITATATTTATTDESKTETKDNKEDIKILIASPSSTQVPQETTTAKASAYSGTHSRTASPDRAGKRVIEQTDKAHPRSLNLSATLNQRNTVPEEELNPKTIASQNAQRKRKMEHDGSLFQAAGIGTLLQQPGDCAPTMSLSWKRVKGFISSEQNGTEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN foxa2_mouse Hepatocyte nuclear factor 3-beta Foxa2 Hnf3b Tcf-3b Tcf3b Forkhead box protein A2 protein protein mouse Mus musculus Mouse MLGAVKMEGHEPSDWSSYYAEPEGYSSVSNMNAGLGMNGMNTYMSMSAAAMGGGSGNMSAGSMNMSSYVGAGMSPSLAGMSPGAGAMAGMSGSAGAAGVAGMGPHLSPSLSPLGGQAAGAMGGLAPYANMNSMSPMYGQAGLSRARDPKTYRRSYTHAKPPYSYISLITMAIQQSPNKMLTLSEIYQWIMDLFPFYRQNQQRWQNSIRHSLSFNDCFLKVPRSPDKPGKGSFWTLHPDSGNMFENGCYLRRQKRFKCEKQLALKEAAGAASSGGKKTAPGSQASQAQLGEAAGSASETPAGTESPHSSASPCQEHKRGGLSELKGAPASALSPPEPAPSPGQQQQAAAHLLGPPHHPGLPPEAHLKPEHHYAFNHPFSINNLMSSEQQHHHSHHHHQPHKMDLKAYEQVMHYPGGYGSPMPGSLAMGPVTNKAGLDASPLAADTSYYQGVYSRPIMNSS arnt_mouse Aryl hydrocarbon receptor nuclear translocator Arnt Dioxin receptor, nuclear translocator Hypoxia-inducible factor 1-beta protein protein mouse Mus musculus Mouse MAATTANPEMTSDVPSLGPTIASGNPGPGIQGGGAVVQRAIKRRSGLDFDDEVEVNTKFLRCDDDQMCNDKERFARSDDEQSSADKERLARENHSEIERRRRNKMTAYITELSDMVPTCSALARKPDKLTILRMAVSHMKSLRGTGNTSTDGSYKPSFLTDQELKHLILEAADGFLFIVSCETGRVVYVSDSVTPVLNQPQSEWFGSTLYDQVHPDDVDKLREQLSTSENALTGRVLDLKTGTVKKEGQQSSMRMCMGSRRSFICRMRCGTSSVDPVSMNRLSFLRNRCRNGLGSVKEGEPHFVVVHCTGYIKAWPPAGVSLPDDDPEAGQGSKFCLVAIGRLQVTSSPNCTDMSNICQPTEFISRHNIEGIFTFVDHRCVATVGYQPQELLGKNIVEFCHPEDQQLLRDSFQQVVKLKGQVLSVMFRFRSKTREWLWMRTSSFTFQNPYSDEIEYIICTNTNVKNSSQEPRPTLSNTIPRSQLGPTANLSLEMGTGQLPSRQQQQQHTELDMVPGRDGLASYNHSQVSVQPVASAGSEHSKPLEKSEGLFAQDRDPRFPEIYPSITADQSKGISSSTVPATQQLFSQGSSFPPNPRPAENFRNSGLTPPVTIVQPSSSAGQILAQISRHSNPAQGSAPTWTSSSRPGFAAQQVPTQATAKTRSSQFGVNNFQTSSSFSAMSLPGAPTASSGTAAYPALPNRGSNFPPETGQTTGQFQARTAEGVGVWPQWQGQQPHHRSSSSEQHVQQTQAQAPSQPEVFQEMLSMLGDQSNTYNNEEFPDLTMFPPFSE q8bj14_mouse Ep300 protein protein mouse Mus musculus Mouse MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDISQLQTSLGIVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQAMASQAQQNSPGLSLINSMVKSPMAQTGLTSPNMGIGSSGPNQGPTQSPAGMMNSPVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRPNMQYPNAGMGNAGSLLTEPLQQGSPQMGGQPGLRGPQPLKMGMMNNPSPYGSPYTQNSGQQIGASGLGLQIQTKTVLPNNLSPFAMDKKAVPGGGMPSMGQQPTPSVQQPGLVTPVAAGMGSGAHTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQSILTGAPVGLGNPSSLGVGQQSTPSLSTVSQIDPSSIERAYAALGLPYQVNQIPPQPQVQAKNQQSQPSGQSPQGMRSVNNMSASPMGVNGGVGVQTPNLLSDSMLHSTINSQNPMMSENAGVASLGPLPTAAQPSSTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAPGMGPVPMNTGSNMGQQPTGMTTNGPVPDPSMIRGSVPNHMMPRMTPQPGNV vegfa_mouse_gene Mouse VEGFA gene gene gene mouse Mus musculus Mouse kdm3a_mouse_gene Mouse Jmjd1a/2a gene gene gene mouse Mus musculus Mouse 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 vegfa_human_gene Human VEGFA gene gene gene human Homo sapiens Human hes6_human_gene Human HES6 gene gene gene human Homo sapiens Human kdm3a_human_gene Human Jmjd1a/2a gene gene gene human Homo sapiens Human foxa2_rat Hepatocyte nuclear factor 3-beta Foxa2 Hnf3b Tcf-3b Tcf3b Forkhead box protein A2 protein protein rat Rattus norvegicus (Rat) Rat MLGAVKMEGHEPSDWSSYYAEPEGYSSVSNMNASLGMNGMNTYMSMSAAAMGSGSGNMSAGSMNMSSYVGAGMSPSLAGMSPGAGAMAGMSGSAGAAGVAGMGPHLSPSLSPLGGQAAGAMGGLAPYANMNSMSPMYGQAGLSRARDPKTYRRSYTHAKPPYSYISLITMAIQQSPNKMLTLSEIYQWIMDLFPFYRQNQQRWQNSIRHSLSFNDFLKVPRAPDKPGKGSFWTLHPDSGNMFENGCYLRRQKRFKCENELALKEAAGAGSGGGKKTAPGTQASQVQLGEAAGSASETPAGTESPHSSASPCQEHKRGGLSELKGTPASALSPPEPAPSPGQQQQAAAHLLGPPHHPGLPPEAHLKPEHHYAFNHPFSINNLMSSEQQHHHSHHHHQPHKMDLKTYEQVMHYPGGYGSPMPGSLAMGPVTNKAGLDASPLAADTSYYQGVYSRPIMNSS hif1a-hes6 972037 n/a 972042 unspecified role unspecified role prey prey n/a 972043 unspecified role unspecified role bait bait association association false false false 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 972037 n/a 972044 unspecified role unspecified role bait bait n/a 972042 unspecified role unspecified role prey prey association association false false false 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 972037 n/a 972045 unspecified role unspecified role bait bait n/a 972042 unspecified role unspecified role prey prey association association false false false 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 972037 n/a 972046 unspecified role unspecified role bait bait n/a 972042 unspecified role unspecified role prey prey association association false false false 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. p300-hes6-2 972037 n/a 972042 unspecified role unspecified role prey prey n/a 972046 unspecified role unspecified role bait bait association association false false false 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-vegfa 972037 n/a 972047 unspecified role unspecified role prey prey n/a 972046 unspecified role unspecified role bait bait association association false false false 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 972037 n/a 972046 unspecified role unspecified role bait bait n/a 972048 unspecified role unspecified role prey prey association association false false false 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. his-foxa2-hif1a 972038 n/a 972044 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 false ecobb Escherichia coli (strain B / BL21) n/a 972043 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 false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 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. hifa1-foxa2 972038 n/a 972044 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 false n terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-tad domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false in vitro In vitro n/a 972043 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 false in vitro In vitro physical association physical association false false false 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. hif1a-foxa2-hif1b 972038 n/a 972045 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 false in vitro In vitro n/a 972044 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 false n terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro n/a 972043 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 false in vitro In vitro physical association physical association false false false Fig 3K. 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. p300-hif1a-foxa2 972038 n/a 972049 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 false in vitro In vitro n/a 972044 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 false n terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro n/a 972043 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 false 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 false in vitro In vitro association association false false false 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. p300-hif531-822 972038 n/a 972049 unspecified role unspecified role bait bait in vitro In vitro n/a 972043 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 false c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false in vitro In vitro physical association physical association false false false 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. hif1a-foxa2-arnt 972039 n/a 972045 unspecified role unspecified role prey prey n/a 972043 unspecified role unspecified role bait bait n/a 972044 unspecified role unspecified role prey prey association association false false false 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 972039 n/a 972045 unspecified role unspecified role bait bait n/a 972044 unspecified role unspecified role prey prey n/a 972043 unspecified role unspecified role prey prey association association false false false Endogenous FoxA2 was coprecipitated with endogenous HIF-1a (Figure 3F) or HIF-1b (Figure 3G) in TRAMP-C cells maintained under hypoxia. Fig 3G. hif1a-foxa2-arnt-1 972039 n/a 972045 unspecified role unspecified role prey prey n/a 972043 unspecified role unspecified role bait bait n/a 972044 unspecified role unspecified role prey prey association association false false false 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. p300-vegfa-1 972040 n/a 972050 unspecified role unspecified role prey prey n/a 972049 unspecified role unspecified role bait bait association association false false false 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 972040 n/a 972049 unspecified role unspecified role bait bait n/a 972051 unspecified role unspecified role prey prey association association false false false 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 972040 n/a 972049 unspecified role unspecified role bait bait n/a 972052 unspecified role unspecified role prey prey association association false false false 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. hif1a-foxa2 972041 n/a 972053 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 false n/a 972043 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 false physical association physical association false false false HA-FoxA2, but not HA-FoxA1, was identified as an HIF-associated protein. Fig 3E.