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/ bullock-2016-5 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-6 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. jclifton@med.usyd.edu.au Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-1 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-2 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-3 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-4 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-7 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-8 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation bullock-2016-9 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 Oncotarget 2016 Only protein-protein interactions imex curation imex curation 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 tert_human_gene Telomerase reverse transcriptase TERT EST2 TCS1 TRT gene gene human Homo sapiens Human elk1_human ETS domain-containing protein Elk-1 ELK1 protein protein human Homo sapiens Human MDPSVTLWQFLLQLLREQGNGHIISWTSRDGGEFKLVDAEEVARLWGLRKNKTNMNYDKLSRALRYYYDKNIIRKVSGQKFVYKFVSYPEVAGCSTEDCPPQPEVSVTSTMPNVAPAAIHAAPGDTVSGKPGTPKGAGMAGPGGLARSSRNEYMRSGLYSTFTIQSLQPQPPPHPRPAVVLPSAAPAGAAAPPSGSRSTSPSPLEACLEAEEAGLPLQVILTPPEAPNLKSEELNVEPGLGRALPPEVKVEGPKEELEVAGERGFVPETTKAEPEVPPQEGVPARLPAVVMDTAGQAGGHAASSPEISQPQKGRKPRDLELPLSPSLLGGPGPERTPGSGSGSGLQAPGPALTPSLLPTHTLTPVLLTPSSLPPSIHFWSTLSPIAPRSPAKLSFQFPSSGSAQVHIPSISVDGLSTPVVLSPGPQKP pert_human_gene Thyroid peroxidase TPO gene gene human Homo sapiens Human thyroglobulin_rat_k_region_promoter_probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid rat Rattus norvegicus (Rat) Rat GAGGGAGTTCCTGTGACTAGCAGAGAAAACAAAGTGAGCCAC tert_human_promoter_probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human CCCCGCCCCGTCCCGACCCCTCCCGGGTCCCCGGCCCAGCCCCCTCCGGGCCCTCCCAGCCCCTCCCCTTCCTTTCCGCGGCCCC rel_human Proto-oncogene c-Rel REL protein protein human Homo sapiens Human MASGAYNPYIEIIEQPRQRGMRFRYKCEGRSAGSIPGEHSTDNNRTYPSIQIMNYYGKGKVRITLVTKNDPYKPHPHDLVGKDCRDGYYEAEFGQERRPLFFQNLGIRCVKKKEVKEAIITRIKAGINPFNVPEKQLNDIEDCDLNVVRLCFQVFLPDEHGNLTTALPPVVSNPIYDNRAPNTAELRICRVNKNCGSVRGGDEIFLLCDKVQKDDIEVRFVLNDWEAKGIFSQADVHRQVAIVFKTPPYCKAITEPVTVKMQLRRPSDQEVSESMDFRYLPDEKDTYGNKAKKQKTTLLFQKLCQDHVETGFRHVDQDGLELLTSGDPPTLASQSAGITVNFPERPRPGLLGSIGEGRYFKKEPNLFSHDAVVREMPTGVSSQAESYYPSPGPISSGLSHHASMAPLPSSSWSSVAHPTPRSGNTNPLSSFSTRTLPSNSQGIPPFLRIPVGNDLNASNACIYNNADDIVGMEASSMPSADLYGISDPNMLSNCSVNMMTTSSDSMGETDNPRLLSMNLENPSCNSVLDPRDLRQLHQMSSSSMSAGANSNTTVFVSQSDAFEGSDFSCADNSMINESGPSNSTNPNSHGFVQDSQYSGIGSMQNEQLSDSFPYEFFQV foxe1-tert-1 3559331 N/A 3559340 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep N/A 3559341 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 false physical association physical association false false false 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 3559331 N/A 3559341 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 false N/A 3559342 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physical association physical association false false false 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-3 3559331 N/A 3559340 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep N/A 3559343 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 false physical association physical association false false false Figure 5A foxe1-tpo-1 3559331 N/A 3559343 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 false N/A 3559342 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physical association physical association false false false Figure 5A thyroglobulin_rat_k_region_promoter_probe-foxe1-1 3559332 N/A 3559340 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 false 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. N/A 3559344 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Figure 2B LHS Positive control tert_human_promoter_probe-foxe1-1 3559332 N/A 3559342 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 false 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. N/A 3559345 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 false 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 false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false synthetic construct synthetic construct physical association physical association false false false Figure 2B RHS-1, Supplementary Figure 2 tert_human_promoter_probe-foxe1-2 3559332 N/A 3559340 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 false O00358:p.Ala65Val mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false 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. N/A 3559342 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 false 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. N/A 3559345 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false g.44C>T mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false g.22C>T mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false synthetic construct synthetic construct association association false false false 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 3559332 N/A 3559340 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. N/A 3559342 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 false 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. N/A 3559345 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 false g.22C>T mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false synthetic construct synthetic construct association association false false false Supplementary Figure 2 foxe1-elk1-1 3559333 N/A 3559340 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 false 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 N/A 3559342 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 false false false Figure 1B Other visible spots are signals for FOXF2, FOXD1 and FOXI1 binding sites, and are likely false-positives. foxe1-elk1-2 3559333 N/A 3559340 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 false 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 N/A 3559346 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 false false false Figure 1B foxe1-elk1-3 3559334 N/A 3559340 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 false 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 false N/A 3559342 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 false physical association physical association false false false Figure 1C, 2C foxe1-elk1-4 3559334 N/A 3559340 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 false N/A 3559342 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 false physical association physical association false false false Figure 1F All ELK1 truncations tested could be co-precipitated with FOXE1-Flag. 2345rwet-1 3559335 N/A 3559340 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3559342 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 1D elk1-foxe1-1 3559336 N/A 3559340 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed N/A 3559342 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association false false false Figure 1E foxe1-elk1-5 3559337 N/A 3559340 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 false N/A 3559342 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false 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 false physical association physical association false false false 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. foxe1-elk1-6 3559338 N/A 3559340 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed N/A 3559342 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association false false false Figure 4B U0126 -a MEK inhibitor foxe1-elk1-7 3559339 N/A 3559340 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 false 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 false p.[Ala164_Ala168del] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false O00358:p.Ala179_Ile180insAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false N/A 3559342 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association false false false Supplementary Figure 4 The FOXE1 Uniprot reference sequence used here has a Alanine polyalanine tract of 16 Alanines in length. Increasing the polyalanine tract length had an inhibitory effect upon Gal4-FOXE1 and VP16-ELK1 stimulated promoter activity.