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/ hinz-2018-2 crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag visualisation tag visualisation Hinz S., Navarro G., Borroto-Escuela D., Seibt BF., Ammon YC., de Filippo E., Danish A., Lacher SK., Červinková B., Rafehi M., Fuxe K., Schiedel AC., Franco R., Müller CE. Dopamine D2 receptor assumed to be of Human origin based on previous work by donor, Franco R.: pmid:23637801, 23296780. christa.mueller@uni-bonn.de Oncotarget 2018 Only protein-protein interactions imex curation imex curation hinz-2018-1 crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Hinz S., Navarro G., Borroto-Escuela D., Seibt BF., Ammon YC., de Filippo E., Danish A., Lacher SK., Červinková B., Rafehi M., Fuxe K., Schiedel AC., Franco R., Müller CE. christa.mueller@uni-bonn.de Oncotarget 2018 Only protein-protein interactions imex curation imex curation hinz-2018-4 crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Hinz S., Navarro G., Borroto-Escuela D., Seibt BF., Ammon YC., de Filippo E., Danish A., Lacher SK., Červinková B., Rafehi M., Fuxe K., Schiedel AC., Franco R., Müller CE. christa.mueller@uni-bonn.de Oncotarget 2018 Only protein-protein interactions imex curation imex curation hinz-2018-3 rat-brain-hippocampus Hippocampus of rat brain rat-brain-hippocampus Hippocampus of rat brain pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Hinz S., Navarro G., Borroto-Escuela D., Seibt BF., Ammon YC., de Filippo E., Danish A., Lacher SK., Červinková B., Rafehi M., Fuxe K., Schiedel AC., Franco R., Müller CE. christa.mueller@uni-bonn.de Oncotarget 2018 Only protein-protein interactions imex curation imex curation hinz-2018-5 crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Hinz S., Navarro G., Borroto-Escuela D., Seibt BF., Ammon YC., de Filippo E., Danish A., Lacher SK., Červinková B., Rafehi M., Fuxe K., Schiedel AC., Franco R., Müller CE. christa.mueller@uni-bonn.de Oncotarget 2018 Only protein-protein interactions imex curation imex curation aa2br_human Adenosine receptor A2b ADORA2B protein protein human Homo sapiens Human MLLETQDALYVALELVIAALSVAGNVLVCAAVGTANTLQTPTNYFLVSLAAADVAVGLFAIPFAITISLGFCTDFYGCLFLACFVLVLTQSSIFSLLAVAVDRYLAICVPLRYKSLVTGTRARGVIAVLWVLAFGIGLTPFLGWNSKDSATNNCTEPWDGTTNESCCLVKCLFENVVPMSYMVYFNFFGCVLPPLLIMLVIYIKIFLVACRQLQRTELMDHSRTTLQREIHAAKSLAMIVGIFALCWLPVHAVNCVTLFQPAQGKNKPKWAMNMAILLSHANSVVNPIVYAYRNRDFRYTFHKIISRYLLCQADVKSGNGQAGVQPALGVGL aa2ar_human Adenosine receptor A2a ADORA2A ADORA2 protein protein human Homo sapiens Human MPIMGSSVYITVELAIAVLAILGNVLVCWAVWLNSNLQNVTNYFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFIACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGTRAKGIIAICWVLSFAIGLTPMLGWNNCGQPKEGKNHSQGCGEGQVACLFEDVVPMNYMVYFNFFACVLVPLLLMLGVYLRIFLAARRQLKQMESQPLPGERARSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCPDCSHAPLWLMYLAIVLSHTNSVVNPFIYAYRIREFRQTFRKIIRSHVLRQQEPFKAAGTSARVLAAHGSDGEQVSLRLNGHPPGVWANGSAPHPERRPNGYALGLVSGGSAQESQGNTGLPDVELLSHELKGVCPEPPGLDDPLAQDGAGVS drd2_human D(2) dopamine receptor DRD2 Dopamine D2 receptor protein protein human Homo sapiens Human MDPLNLSWYDDDLERQNWSRPFNGSDGKADRPHYNYYATLLTLLIAVIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMISIVWVLSFTISCPLLFGLNNADQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRRRRKRVNTKRSSRAFRAHLRAPLKGNCTHPEDMKLCTVIMKSNGSFPVNRRRVEAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSTPDSPAKPEKNGHAKDHPKIAKIFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFLKILHC aa2br_rat Adenosine receptor A2b Adora2b protein protein rat Rattus norvegicus (Rat) Rat MQLETQDALYVALELVIAALAVAGNVLVCAAVGASSALQTPTNYFLVSLATADVAVGLFAIPFAITISLGFCTDFHSCLFLACFVLVLTQSSIFSLLAVAVDRYLAIRVPLRYKGLVTGTRARGIIAVLWVLAFGIGLTPFLGWNSKDRATSNCTEPGDGITNKSCCPVKCLFENVVPMSYMVYFNFFGCVLPPLLIMMVIYIKIFMVACKQLQHMELMEHSRTTLQREIHAAKSLAMIVGIFALCWLPVHAINCITLFHPALAKDKPKWVMNVAILLSHANSVVNPIVYAYRNRDFRYSFHRIISRYVLCQTDTKGGSGQAGGQSTFSLSL aa2ar_rat Adenosine receptor A2a Adora2a protein protein rat Rattus norvegicus (Rat) Rat MGSSVYITVELAIAVLAILGNVLVCWAVWINSNLQNVTNFFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFFACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGVRAKGIIAICWVLSFAIGLTPMLGWNNCSQKDGNSTKTCGEGRVTCLFEDVVPMNYMVYYNFFAFVLLPLLLMLAIYLRIFLAARRQLKQMESQPLPGERTRSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCSTCRHAPPWLMYLAIILSHSNSVVNPFIYAYRIREFRQTFRKIIRTHVLRRQEPFQAGGSSAWALAAHSTEGEQVSLRLNGHPLGVWANGSATHSGRRPNGYTLGLGGGGSAQGSPRDVELPTQERQEGQEHPGLRGHLVQARVGASSWSSEFAPS adora2b-adora2a-1 3575584 N/A 3575589 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed 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 3575590 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed 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 2 The BRET signal after 60 min of treatment with agonists (adenosine, nonselective; NECA, nonselective; CGS-21680, A2A-selective; BAY60–6583, A2B-selective) or the A2B antagonist PSB-603 was similar to that obtained in the absence of ligands. adora2b-adora2a-2 3575584 N/A 3575590 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed 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 3575591 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed 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 Figure 2 Positive control adora2b-adora2a-3 3575584 N/A 3575589 competitor competitor competitor unspecified role unspecified role over-expressed over expressed level over-expressed N/A 3575589 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed 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 3575590 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed 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 2i adora2a-adora2b-1 3575585 N/A 3575589 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed C-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3575590 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure I-2 adora2a-9 3575585 N/A 3575590 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed 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 3575590 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed C-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag 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 I adora2a-11 3575586 N/A 3575590 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus nyfp nYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3575590 unspecified role unspecified role neutral component neutral component 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 C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Supplementary Figure 2A adora2a-adora2a-6 3575586 N/A 3575589 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus nyfp nYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3575590 unspecified role unspecified role neutral component neutral component 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 C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Supplementary Figure 2B adora2b-adora2a-4 3575587 N/A 3575592 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3575593 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 3, Supplementary figure 4/5 PLA clusters were seen in the CA3 region of the dorsal hippocampus and in the polymorphic layer of the dentate gyrus. The CA1 region showed a similar distribution pattern (as compared with CA3), with high dot/cluster densities within the pyramidal cell layer. However, the diameter range of the clusters, were reduced in this CA1 region versus CA3. adora2a-adora2b-2 3575588 N/A 3575589 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed N/A 3575590 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed proximity proximity close-contact colocalization neighbourhood interaction false false false Supplementary Figure 3