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/ siddiquee-2013-1 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. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Siddiquee K., Hampton J., McAnally D., May L., Smith L. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation siddiquee-2013-2 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. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Siddiquee K., Hampton J., McAnally D., May L., Smith L. Cross-linking with DTSSP(3,3'-dithiobis-sulfosucciniimidyl propionate) was carried out before or after immunoprecipitation. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation siddiquee-2013-4 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. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag visualisation tag visualisation Siddiquee K., Hampton J., McAnally D., May L., Smith L. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation siddiquee-2013-7 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 Siddiquee K., Hampton J., McAnally D., May L., Smith L. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation siddiquee-2013-5 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 anti tag western anti tag western blot anti tag western Siddiquee K., Hampton J., McAnally D., May L., Smith L. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation siddiquee-2013-6 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. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Siddiquee K., Hampton J., McAnally D., May L., Smith L. Cross-linking with DTSSP(3,3'-dithiobis-sulfosucciniimidyl propionate) was carried out before or after immunoprecipitation. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation siddiquee-2013-3 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. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag visualisation tag visualisation Siddiquee K., Hampton J., McAnally D., May L., Smith L. The authors have stated that constructs used in figure 2 are all of human origin. lhsmith@sanfordburnham.org British journal of pharmacology 2013 Only protein-protein interactions imex curation imex curation apj_human Apelin receptor APLNR AGTRL1 APJ Angiotensin receptor-like 1 G-protein coupled receptor APJ G-protein coupled receptor HG11 protein protein human Homo sapiens Human MEEGGDFDNYYGADNQSECEYTDWKSSGALIPAIYMLVFLLGTTGNGLVLWTVFRSSREKRRSADIFIASLAVADLTFVVTLPLWATYTYRDYDWPFGTFFCKLSSYLIFVNMYASVFCLTGLSFDRYLAIVRPVANARLRLRVSGAVATAVLWVLAALLAMPVMVLRTTGDLENTTKVQCYMDYSMVATVSSEWAWEVGLGVSSTTVGFVVPFTIMLTCYFFIAQTIAGHFRKERIEGLRKRRRLLSIIVVLVVTFALCWMPYHLVKTLYMLGSLLHWPCDFDLFLMNIFPYCTCISYVNSCLNPFLYAFFDPRFRQACTSMLCCGQSRCAGTSHSSSGEKSASYSSGHSQGPGPNMGKGGEQMHEKSIPYSQETLVVD agtr1_human Type-1 angiotensin II receptor AGTR1 AGTR1A AGTR1B AT2R1 AT2R1B AT1AR AT1BR Angiotensin II type-1 receptor protein protein human Homo sapiens Human MILNSSTEDGIKRIQDDCPKAGRHNYIFVMIPTLYSIIFVVGIFGNSLVVIVIYFYMKLKTVASVFLLNLALADLCFLLTLPLWAVYTAMEYRWPFGNYLCKIASASVSFNLYASVFLLTCLSIDRYLAIVHPMKSRLRRTMLVAKVTCIIIWLLAGLASLPAIIHRNVFFIENTNITVCAFHYESQNSTLPIGLGLTKNILGFLFPFLIILTSYTLIWKALKKAYEIQKNKPRNDDIFKIIMAIVLFFFFSWIPHQIFTFLDVLIQLGIIRDCRIADIVDTAMPITICIAYFNNCLNPLFYGFLGKKFKRYFLQLLKYIPPKAKSHSNLSTKMSTLSYRPSDNVSSSTKKPAPCFEVE agtr1-aplnr-1 3575106 N/A 3575113 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3575114 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 1A, Lane 1 agtr1-aplnr-3 3575106 N/A 3575113 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3575114 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 1B, Lane 1 agtr1-aplnr-2 3575107 N/A 3575113 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3575114 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 1A, Lane 2 +3 agtr1-aplnr-4 3575107 N/A 3575113 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3575114 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 1B, Lane 2 +3 aplnr-agtr1-2 3575108 N/A 3575113 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept physiological level physiological level endogenous level endogenous endogenous endogenous level C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3575114 unspecified role unspecified role luminescence donor luminescence donor physiological level physiological level endogenous level endogenous endogenous endogenous level C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Supplementary figure 4 aplnr-agtr1-3 3575108 N/A 3575113 unspecified role unspecified role luminescence donor luminescence donor physiological level physiological level endogenous level endogenous endogenous endogenous level C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3575114 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept physiological level physiological level endogenous level endogenous endogenous endogenous level C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Supplementary figure 4 aplnr-agtr1-5 3575109 N/A 3575113 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3575114 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 Figure 2C Apelin-13, alone, and in presence of Angiotensin II. agtr1-aplnr-8 3575109 N/A 3575113 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3575114 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 Figure 2D Apelin-13, alone, and in presence of Angiotensin II. aplnr-agtr1-6 3575110 N/A 3575113 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3575114 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 Figure 2A agtr1-aplnr-9 3575110 N/A 3575113 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3575114 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 Figure 2A agtr1-aplnr-5 3575111 N/A 3575114 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 N/A 3575113 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2B aplnr-agtr1-7 3575111 N/A 3575113 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3575114 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 Figure 2B aplnr-agtr1-1 3575112 N/A 3575113 unspecified role unspecified role luminescence donor luminescence donor physiological level physiological level endogenous level endogenous endogenous endogenous level C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3575114 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept physiological level physiological level endogenous level endogenous endogenous endogenous level C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 3, Supplementary figure 3 Apelin -13 peptide