MRAP2 inhibits PKCα recruitment to GHSR1a.A, Western blot detection of immunoprecipitated 3HA-GHSR1a from cells transfected with 3HA-GHSR1a, loaded with 32P, and treated with vehicle, ghrelin, ghrelin and Go6983, ghrelin and Compound 101, or ghrelin and both Go6983 and Compound 101. B, phosphoimaging detecting 32P incorporation in GHSR1a in the same samples shown in (A). C, densitometry analysis of GHSR1a phosphorylation in samples from cells treated with vehicle, ghrelin, ghrelin and Go6983, ghrelin and Compound 101, or ghrelin and both Go6983 and Compound 101. D, ghrelin stimulated β-arrestin2 recruitment to GHSR1a with and without MRAP2 in the presence or absence of Go6983. Arrow indicates ghrelin injection time. E, ghrelin stimulated β-arrestin1 recruitment to GHSR1a with and without MRAP2 in the presence or absence of Go6983. F, ghrelin stimulated β-arrestin2 recruitment to GHSR1a with and without MRAP2 in the presence or absence of Compound 101 and Go6983. G, ghrelin stimulated PKCα recruitment to GHSR1a with and without MRAP2. Data are the mean ± SEM of three independent experiments. ∗∗∗∗p < 0.0001. GHSR1a, growth hormone secretagogue receptor 1a; HA, hemagglutinin.