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/ Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Oncotarget 2018 imex curation 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 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. HAd great difficulty in finding RLUC tag locations:"pRluc-N vectors contain a multiple cloning site (MCS) located upstream of the Renilla Luciferase gene (Rluc)" 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 Accepted 2019-JUN-07 AT 12:27 BST AT 12:27 BST by LIVIAP Oncotarget 2018 Only protein-protein interactions imex curation imex curation altered; I have deleted EBI-21408114 as I cannot find it either. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Oncotarget 2018 imex curation 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 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2019-JUN-07 AT 12:27 BST AT 12:27 BST by LIVIAP Oncotarget 2018 Only protein-protein interactions imex curation imex curation altered; I have deleted EBI-21408114 as I cannot find it either. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Oncotarget 2018 imex curation 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 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2019-JUN-07 AT 12:27 BST AT 12:27 BST by LIVIAP Oncotarget 2018 Only protein-protein interactions imex curation imex curation altered; I have deleted EBI-21408114 as I cannot find it either. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Oncotarget 2018 imex curation 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 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. cannot include Jun/Fos positive control as no species given and one of the Refs given = 63, does not exist in supplement or main paper - last ref number is 62. The other reference is Supplementary 1(donor = Kerppola) - which does not mention the species or any mutation info. christa.mueller@uni-bonn.de Accepted 2019-JUN-07 AT 12:27 BST AT 12:27 BST by LIVIAP Oncotarget 2018 Only protein-protein interactions imex curation imex curation altered; I have deleted EBI-21408114 as I cannot find it either. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors. Oncotarget 2018 imex curation 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 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2019-JUN-07 AT 12:27 BST AT 12:27 BST by LIVIAP Oncotarget 2018 Only protein-protein interactions imex curation imex curation altered; I have deleted EBI-21408114 as I cannot find it either. aa2ar_human Adenosine receptor A2a ADORA2A ADORA2 protein protein human Homo sapiens Human MPIMGSSVYITVELAIAVLAILGNVLVCWAVWLNSNLQNVTNYFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFIACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGTRAKGIIAICWVLSFAIGLTPMLGWNNCGQPKEGKNHSQGCGEGQVACLFEDVVPMNYMVYFNFFACVLVPLLLMLGVYLRIFLAARRQLKQMESQPLPGERARSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCPDCSHAPLWLMYLAIVLSHTNSVVNPFIYAYRIREFRQTFRKIIRSHVLRQQEPFKAAGTSARVLAAHGSDGEQVSLRLNGHPPGVWANGSAPHPERRPNGYALGLVSGGSAQESQGNTGLPDVELLSHELKGVCPEPPGLDDPLAQDGAGVS 9438E9D64A6BE61B aa2br_rat Adenosine receptor A2b Adora2b protein protein rat Rattus norvegicus (Rat) Rat MQLETQDALYVALELVIAALAVAGNVLVCAAVGASSALQTPTNYFLVSLATADVAVGLFAIPFAITISLGFCTDFHSCLFLACFVLVLTQSSIFSLLAVAVDRYLAIRVPLRYKGLVTGTRARGIIAVLWVLAFGIGLTPFLGWNSKDRATSNCTEPGDGITNKSCCPVKCLFENVVPMSYMVYFNFFGCVLPPLLIMMVIYIKIFMVACKQLQHMELMEHSRTTLQREIHAAKSLAMIVGIFALCWLPVHAINCITLFHPALAKDKPKWVMNVAILLSHANSVVNPIVYAYRNRDFRYSFHRIISRYVLCQTDTKGGSGQAGGQSTFSLSL F0ABAB5CCACA8858 drd2_human D(2) dopamine receptor DRD2 Dopamine D2 receptor protein protein human Homo sapiens Human MDPLNLSWYDDDLERQNWSRPFNGSDGKADRPHYNYYATLLTLLIAVIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMISIVWVLSFTISCPLLFGLNNADQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRRRRKRVNTKRSSRAFRAHLRAPLKGNCTHPEDMKLCTVIMKSNGSFPVNRRRVEAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSTPDSPAKPEKNGHAKDHPKIAKIFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFLKILHC 9BF8EA36C988A2E2 aa2ar_rat Adenosine receptor A2a Adora2a protein protein rat Rattus norvegicus (Rat) Rat MGSSVYITVELAIAVLAILGNVLVCWAVWINSNLQNVTNFFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFFACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGVRAKGIIAICWVLSFAIGLTPMLGWNNCSQKDGNSTKTCGEGRVTCLFEDVVPMNYMVYYNFFAFVLLPLLLMLAIYLRIFLAARRQLKQMESQPLPGERTRSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCSTCRHAPPWLMYLAIILSHSNSVVNPFIYAYRIREFRQTFRKIIRTHVLRRQEPFQAGGSSAWALAAHSTEGEQVSLRLNGHPLGVWANGSATHSGRRPNGYTLGLGGGGSAQGSPRDVELPTQERQEGQEHPGLRGHLVQARVGASSWSSEFAPS D87B256CD2336BAC aa2br_human Adenosine receptor A2b ADORA2B protein protein human Homo sapiens Human MLLETQDALYVALELVIAALSVAGNVLVCAAVGTANTLQTPTNYFLVSLAAADVAVGLFAIPFAITISLGFCTDFYGCLFLACFVLVLTQSSIFSLLAVAVDRYLAICVPLRYKSLVTGTRARGVIAVLWVLAFGIGLTPFLGWNSKDSATNNCTEPWDGTTNESCCLVKCLFENVVPMSYMVYFNFFGCVLPPLLIMLVIYIKIFLVACRQLQRTELMDHSRTTLQREIHAAKSLAMIVGIFALCWLPVHAVNCVTLFQPAQGKNKPKWAMNMAILLSHANSVVNPIVYAYRNRDFRYTFHKIISRYLLCQADVKSGNGQAGVQPALGVGL 38AD088E2439C7FA adora2a-adora2b-1 2 10 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 6 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain proximity proximity close-contact colocalization neighbourhood interaction Figure I-2 adora2a-9 2 6 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 6 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 proximity proximity close-contact colocalization neighbourhood interaction Figure I adora2b-adora2a-1 1 10 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 6 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 proximity proximity close-contact colocalization neighbourhood interaction 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 1 6 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 8 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 2 Positive control adora2b-adora2a-3 1 10 competitor competitor competitor unspecified role unspecified role over-expressed over expressed level over-expressed 10 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 6 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 2i adora2b-adora2a-4 3 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 9 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 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-11 4 6 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 6 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 C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Supplementary Figure 2A adora2a-adora2a-6 4 10 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 6 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 C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Supplementary Figure 2B adora2a-adora2b-2 5 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed proximity proximity close-contact colocalization neighbourhood interaction Supplementary Figure 3