Extended Data Fig. 7: GPCR-induced βarr conformational change for the NC-site βarr mutant.
From: Membrane-domain compartmentalization of active GPCRs by β-arrestins through PtdIns(4,5)P2 binding
(a) Concentration-response curves illustrate the change in the endpoint of Ib30 interaction for WT, C-3Q and NC-4Q for both βarr1 and βarr2 in the V2R. Symbols and error bars represent mean and SEM, respectively, 3 independent experiments with each performed in duplicate. (b) Normalized efficacy of endpoint of Ib30 interaction. For each experiment, these data were normalized to WT response. (c) Normalization of the Ib30 assay to the level of membrane recruitment. For each experiment, data were normalized to WT response. Bars and error bars are mean and SEM, respectively, of three independent experiments with each performed duplicate. (d) Concentration-response curves of the HiBiT-based V2R internalization assay. HiBiT-V2R and EGFP-βarr2 (WT, C-3Q or NC-4Q) were co-expressed in ∆βarr1/2 HEK293 cells and AVP-induced HiBiT-V2R internalization was monitored. Symbols and error bars represent mean and SEM, respectively, three independent experiments with each performed in duplicate. For the statistical analyses, data were analyzed by one-way ANOVA followed by the Dunnett’s test for multiple comparison analysis. ns, not significantly different between the groups. *P < 0.05 vs. WT.
