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Image Search Results
Journal: Nature Communications
Article Title: G z ESTY as an optimized cell-based assay for initial steps in GPCR deorphanization
doi: 10.1038/s41467-025-59850-8
Figure Lengend Snippet: A Real-time analysis of ligand-mediated activation of a battery of G i/o/z -PCRs (red) and G s -PCR (green) in cells co-transfected with indicated receptors and the 3-in-one G z ESTY plasmid in the presence of 50 µM IBMX. Cells transfected only with the 3-in-one plasmid served as control (white). The following agonists were applied at 4 min: 10 µM dopamine (D2R, D1R), 100 µM clonidine (ADRA2A, ADRA2B, ADRA2C), 10 µM GABA (GABABR), 10 µM DAMGO (MOR), 1 µM SNC-80 (DOR), 10 µM salvinorin A (KOR), 10 µM human galanin (1–30) (GAL1R), 100 µM serotonin (5-HT1B), 10 µM human neuropeptide Y (13–36) (NPYR1), 10 µM lysophosphatidic acid (LPAR2), 10 µM 2-arachidonoyl glycerol (CB1R, CB2R), 10 µM N-formyl-met-leu-phe (FPR1), 1 mM isobutyric acid (FFAR3), 10 µM somatostatin 14 (SSTR1), 10 µM neuropeptide FF (NPFFR1), 10 µM SEW2871 (S1PR1), 10 µM histamine (HRH3), 10 µM quinpirole (D3R), 10 µM TFLLR (PAR1), 10 µM SLIGKV (PAR2), 10 µM MK-6892 (HCA2R), 1 µM teriparatide PTH (PTH1R), 10 µM AB-MECA (ADORA2B), 10 µM NDP-α-MSH (MC4R), and 10 µM CGRP (CLR). Data are shown as means ± SEM; N = 5 independent transfections. B GPCRs that couple to G s and/or G i/o/z and can potentially be detected by GZESTY are highlighted and include 213 out of 249 total ligand-activated GPCRs (86%) (adapted from GPRCdb.org). C Quantification of agonist-induced activity for 24 G i/o/z -coupled GPCRs. On the right, the response to agonist of the last seven GPCRs is also reported with a different scale. Data are shown as means ± SEM of the fold change obtained; N = 5 independent transfections.
Article Snippet: The following chemicals were purchased: clonidine (Tocris), dopamine (Tocris), GABA (Tocris), serotonin (Tocris), 1-oleoyl lysophosphatidic acid (Tocris), DAMGO (MedChemExpress), TFLLR (MedChemExpress), human PAR-2 (1-6, SLIGKV) (MedChemExpress), human galanin (1–30) (MedChemExpress), IBMX (MedChemExpress),
Techniques: Activation Assay, Battery, Transfection, Plasmid Preparation, Control, Activity Assay