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/ Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. gal4 vp16 complement gal4 vp16 complementation karyoplasmic interaction ion strategy KISS mammalian two hybrid gal4 vp16 complement predetermined predetermined participant predetermined Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-nthy-ori-3.1 nthy-ori-3.1 Normal human primary thyroid follicular epithelial cells nthy-ori-3.1 NThy-ori-3.1 ECACC Catalogue No.: 90011609 Cellosaurus;CVCL_2659 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-sw1736 Thyroid gland undifferentiated (anaplastic) carcinoma sw1736 SW 1736 CVCL_3883 gal4 vp16 complement gal4 vp16 complementation karyoplasmic interaction ion strategy KISS mammalian two hybrid gal4 vp16 complement predetermined predetermined participant predetermined Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro array technology array technology predetermined predetermined participant predetermined Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. Nuclear extracts from NThy-ori-3.1 cells transiently transfected with a 3XFlag-FOXE1 expression plasmid were incubated with a provided set of biotin-labeled double-stranded oligonucleotide probes, which represent a known library of cis-elements. During the incubation step, these probes bind to their corresponding TFs in the nuclear extract. The next step is immunoprecipitation: using an anti-Flag antibody, FOXE1 and associated TFs, along with corresponding cis-elements were isolated. Next, the free cis-elements and nonspecific binding proteins were washed away and the DNA is then separated from protein by washing. Finally, the cis-elements are hybridized to the TranSignalTM Protein/DNA Array membrane. These arrays are spotted with various different consensus sequences depending on the array used. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-thyroid_gland human thyroid gland thyroid_gland ch-ip chromatin immunoprecipitation assay ch-ip q-pcr quantitative pcr q-pcr Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. Thyroid tissue from patients with Graves Disease jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa tag visualisation tag visualisation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. Not sure if an IP alone is enough purification to include these EMSAs - but I see no extraneous bands on the gel, so included, but left as Physical Association rather than "direct". Foxe1 added as "partially purified". jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-thyroid_gland human thyroid gland thyroid_gland anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-sw1736 Thyroid gland undifferentiated (anaplastic) carcinoma sw1736 SW 1736 CVCL_3883 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Thyroid transcription factor FOXE1 interacts with ETS factor ELK1 to co-regulate TERT. Oncotarget 2016 imex curation Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-nthy-ori-3.1 nthy-ori-3.1 Normal human primary thyroid follicular epithelial cells nthy-ori-3.1 NThy-ori-3.1 ECACC Catalogue No.: 90011609 Cellosaurus;CVCL_2659 gal4 vp16 complement gal4 vp16 complementation karyoplasmic interaction ion strategy KISS mammalian two hybrid gal4 vp16 complement predetermined predetermined participant predetermined Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ., Bullock M., Lim G., Li C., Choi IH., Kochhar S., Liddle C., Zhang L., Clifton-Bligh RJ. jclifton@med.usyd.edu.au Accepted 2019-MAY-23 AT 08:48 BST AT 08:48 BST by LIVIAP Oncotarget 2016 Only protein-protein interactions imex curation imex curation EBI-21348979 and EBI-21348921 amalgamated, with a Flag tag. (I do not see there is any interaction without a Flag tag on FOXe1 either.) EBI-21348979 deleted. I have altered the nucleotide mutations on residue 44 and 22 to "mutation decreasing" rather than "disrupting" as the text implies there is NO effect of these mutations on formation of ELK1/FOXE1/probe complex - but I think there is, especially when comparing Fig 2B (big blob) with Supp. fig 2 (faint band)!???; I have altered to Array technology - and altered Roles and I have added "partially purified" as well as "subcellular extract" . However I am not sure we can assign Roles of Bait and Prey here - as no proteins are involved in the array- it is only indirectly shown that these 2 proteins bind. Maybe roles should be "unspecified"? I have added a NEW Exp Mod at the bottom of the box after the "********", but left the OLD one at the top - not sure which one to leave - the NEW one gives greater clarity, but the OLD one is more concise.; done elk1_human ETS domain-containing protein Elk-1 ELK1 protein protein human Homo sapiens Human MDPSVTLWQFLLQLLREQGNGHIISWTSRDGGEFKLVDAEEVARLWGLRKNKTNMNYDKLSRALRYYYDKNIIRKVSGQKFVYKFVSYPEVAGCSTEDCPPQPEVSVTSTMPNVAPAAIHAAPGDTVSGKPGTPKGAGMAGPGGLARSSRNEYMRSGLYSTFTIQSLQPQPPPHPRPAVVLPSAAPAGAAAPPSGSRSTSPSPLEACLEAEEAGLPLQVILTPPEAPNLKSEELNVEPGLGRALPPEVKVEGPKEELEVAGERGFVPETTKAEPEVPPQEGVPARLPAVVMDTAGQAGGHAASSPEISQPQKGRKPRDLELPLSPSLLGGPGPERTPGSGSGSGLQAPGPALTPSLLPTHTLTPVLLTPSSLPPSIHFWSTLSPIAPRSPAKLSFQFPSSGSAQVHIPSISVDGLSTPVVLSPGPQKP 68F71F8ADB9D38CA rel_human Proto-oncogene c-Rel REL protein protein human Homo sapiens Human MASGAYNPYIEIIEQPRQRGMRFRYKCEGRSAGSIPGEHSTDNNRTYPSIQIMNYYGKGKVRITLVTKNDPYKPHPHDLVGKDCRDGYYEAEFGQERRPLFFQNLGIRCVKKKEVKEAIITRIKAGINPFNVPEKQLNDIEDCDLNVVRLCFQVFLPDEHGNLTTALPPVVSNPIYDNRAPNTAELRICRVNKNCGSVRGGDEIFLLCDKVQKDDIEVRFVLNDWEAKGIFSQADVHRQVAIVFKTPPYCKAITEPVTVKMQLRRPSDQEVSESMDFRYLPDEKDTYGNKAKKQKTTLLFQKLCQDHVETGFRHVDQDGLELLTSGDPPTLASQSAGITVNFPERPRPGLLGSIGEGRYFKKEPNLFSHDAVVREMPTGVSSQAESYYPSPGPISSGLSHHASMAPLPSSSWSSVAHPTPRSGNTNPLSSFSTRTLPSNSQGIPPFLRIPVGNDLNASNACIYNNADDIVGMEASSMPSADLYGISDPNMLSNCSVNMMTTSSDSMGETDNPRLLSMNLENPSCNSVLDPRDLRQLHQMSSSSMSAGANSNTTVFVSQSDAFEGSDFSCADNSMINESGPSNSTNPNSHGFVQDSQYSGIGSMQNEQLSDSFPYEFFQV 03E1033BDF7B1B30 thyroglobulin_rat_k_region_promoter_probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid rat Rattus norvegicus (Rat) Rat GAGGGAGTTCCTGTGACTAGCAGAGAAAACAAAGTGAGCCAC pert_human_gene Thyroid peroxidase TPO gene gene human Homo sapiens Human tert_human_gene Telomerase reverse transcriptase TERT EST2 TCS1 TRT gene gene human Homo sapiens Human foxe1_human Forkhead box protein E1 FKHL15 FOXE2 Forkhead box protein E2 Forkhead-related protein FKHL15 HFKH4 HNF-3/fork head-like protein 5 TITF2 TTF2 Thyroid transcription factor 2 FOXE1 protein protein human Homo sapiens Human MTAESGPPPPQPEVLATVKEERGETAAGAGVPGEATGRGAGGRRRKRPLQRGKPPYSYIALIAMAIAHAPERRLTLGGIYKFITERFPFYRDNPKKWQNSIRHNLTLNDCFLKIPREAGRPGKGNYWALDPNAEDMFESGSFLRRRKRFKRSDLSTYPAYMHDAAAAAAAAAAAAAAAAIFPGAVPAARPPYPGAVYAGYAPPSLAAPPPVYYPAASPGPCRVFGLVPERPLSPELGPAPSGPGGSCAFASAGAPATTTGYQPAGCTGARPANPSAYAAAYAGPDGAYPQGAGSAIFAAAGRLAGPASPPAGGSSGGVETTVDFYGRTSPGQFGALGACYNPGGQLGGASAGAYHARHAAAYPGGIDRFVSAM D201EAF05572FCF0 tert_human_promoter_probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human CCCCGCCCCGTCCCGACCCCTCCCGGGTCCCCGGCCCAGCCCCCTCCGGGCCCTCCCAGCCCCTCCCCTTCCTTTCCGCGGCCCC foxe1-elk1-1 4 15 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep partially purified partially purified n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-nthy-ori-3.1 nthy-ori-3.1 Normal human primary thyroid follicular epithelial cells nthy-ori-3.1 NThy-ori-3.1 ECACC Catalogue No.: 90011609 Cellosaurus;CVCL_2659 10 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep partially purified partially purified human-nthy-ori-3.1 nthy-ori-3.1 Normal human primary thyroid follicular epithelial cells nthy-ori-3.1 NThy-ori-3.1 ECACC Catalogue No.: 90011609 Cellosaurus;CVCL_2659 physical association physical association Figure 1B the strongest signal was for the ETS (E26 transformation-specific) factor ELK1. Other visible spots are signals for FOXF2, FOXD1 and FOXI1 binding sites, and are likely false-positives. foxe1-elk1-2 4 15 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep partially purified partially purified n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-nthy-ori-3.1 nthy-ori-3.1 Normal human primary thyroid follicular epithelial cells nthy-ori-3.1 NThy-ori-3.1 ECACC Catalogue No.: 90011609 Cellosaurus;CVCL_2659 11 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep partially purified partially purified human-nthy-ori-3.1 nthy-ori-3.1 Normal human primary thyroid follicular epithelial cells nthy-ori-3.1 NThy-ori-3.1 ECACC Catalogue No.: 90011609 Cellosaurus;CVCL_2659 physical association physical association Figure 1B foxe1-elk1-3 2 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range O00358:p.Ala65Val mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain A V Tue Jun 30 19:08:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala65Val' 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 1C, 2C 2345rwet-1 7 15 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 1D elk1-foxe1-1 1 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association Figure 1E foxe1-elk1-4 2 15 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 1F all ELK1 truncations tested could be co-precipitated with FOXE1-Flag (Figure 1F). All ELK1 truncations tested could be co-precipitated with FOXE1-Flag. foxe1-tert-1 5 15 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep 14 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep promoter_region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 2A There was a 4.6-fold enrichment of FOXE1 in the same region of the TERT promoter (-151 to +11-bp relative to the TSS) that is specifically bound by ELK1. foxe1-tert-2 5 14 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep promoter_region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physical association physical association Figure 2A There was a 4.6-fold enrichment of FOXE1 in the same region of the TERT promoter (-151 to +11-bp relative to the TSS) that is specifically bound by ELK1. thyroglobulin_rat_k_region_promoter_probe-foxe1-1 6 15 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep partially purified partially purified n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 12 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Figure 2B LHS Positive control tert_human_promoter_probe-foxe1-1 6 10 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep partially purified partially purified n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 16 unspecified role unspecified role bait bait g.44C>T mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain C T g.22C>T mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain C T region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined synthetic construct synthetic construct physical association physical association Figure 2B RHS-1, Supplementary Figure 2 tert_human_promoter_probe-foxe1-2 6 15 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep partially purified partially purified n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range O00358:p.Ala65Val mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain A V Tue Jun 30 17:01:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala65Val' human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 10 unspecified role unspecified role prey prey partially purified partially purified n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 16 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined g.44C>T mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain C T g.22C>T mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain C T synthetic construct synthetic construct association association Figure 2B RHS-2,Supplementary Figure 2 The probe was readily bound by ELK1, but unexpectedly was only shifted by FOXE1 when ELK1 was also present. A DNA-binding mutation (p.Ala65Val) in FOXE1 abrogated its interaction with TERT-bound ELK1. tert_human_promoter_probe-elk1-1 6 15 unspecified role unspecified role prey prey partially purified partially purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 10 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep partially purified partially purified n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 16 unspecified role unspecified role bait bait g.44C>T mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C T g.22C>T mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C T region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined synthetic construct synthetic construct association association Supplementary Figure 2 foxe1-elk1-5 8 15 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep P19419:p.[Thr353Xaa;Thr361Xaa;Thr368Xaa;Ser383Xaa;Ser389Xaa;Thr417Xaa] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain T X certain certain sequence position certain certain certain sequence position certain T X certain certain sequence position certain certain certain sequence position certain T X certain certain sequence position certain certain certain sequence position certain S X certain certain sequence position certain certain certain sequence position certain S X certain certain sequence position certain certain certain sequence position certain T X Tue Jun 30 17:05:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr353Xaa;Thr361Xaa;Thr368Xaa;Ser383Xaa;Ser389Xaa;Thr417Xaa]' n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined observed-ptm observed-ptm observed ptm observed-ptm physical association physical association Figure 4A MEK inhibitor PD98059 MEK inhibitor U0126 When all known phospho-accepting Ser/Thr were mutated in ELK, this mutant ELK1-HA showed a significant reduction in its ability co-immunoprecipitate FOXE1-Flag. I would have htought all ELK1 mutations would have been to Alanine(?) but cannot find this stated anywhere, so Xaa/X entered. foxe1-elk1-6 3 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association Figure 4B U0126 -a MEK inhibitor foxe1-tert-3 5 15 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep 13 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep proximal_promoter_region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 5A foxe1-tpo-1 5 13 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep proximal_promoter_region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physical association physical association Figure 5A foxe1-elk1-7 9 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed O00358:p.Ala164_Ala165del mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain AA .. Mon Apr 29 11:32:53 BST 2019 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ala164_Ala165del]' Tue Jun 30 18:35:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala164_Ala165del' p.[Ala164_Ala167del] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain AAAA . p.[Ala164_Ala168del] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain AAAAA . O00358:p.Ala179_Ile180insAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain AI AAI Tue Jun 30 18:43:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala179_Ile180insAla' 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association Supplementary Figure 4 The FOXE1 Uniprot reference sequence used here has a Alanine polyalanine tract of 16 Alanines in length. not sure if the "mutation decreasing" - the 17 Ala's, is really decreasing (statistically significant) - but have entered as such? Increasing the polyalanine tract length had an inhibitory effect upon Gal4-FOXE1 and VP16-ELK1 stimulated promoter activity.