Fig. 4: MRAP2 enhances Gq activation.

A Mean concentration-response curves of the Gq-family selective effector p63RhoGEF-RlucII recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 min, with overexpression of Gq in the presence or absence of MRAP2 or RAMP3 in transiently transfected HEK293-SL cells. MRAP2 or RAMP3 plasmid DNA is co-transfected with the same amount (1:1) with respect to MC4R DNA. Normalized data are expressed as mean ± SEM of three independent experiments. B pEC₅₀ from the p63RhoGEF recruitment BRET experiments for Gq. Results are represented in a bar chart as mean ± SEM, in which individual pEC₅₀ data point from the three individual experiments are shown as overlaid dot plot. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (***: adjusted p-value = 0.0002). C Average concentration-response curves of IP-One assay upon MC4R stimulation using α-MSH, with and without MRAP2 1 + 4 DNA ratio with respect to MC4R, in transiently transfected HEK293T cells. Data are expressed as mean ± standard error of the mean (± SEM) of four independent experiments. D pEC₅₀ values across all the replicates of the IP-One concentration-response curves for the two conditions. Results are represented in a bar chart as mean ± SEM, in which individual pEC₅₀ data point from the four individual experiments are shown as overlaid dot plot. Statistical analysis was performed using an unpaired two-tailed t-test (**: adjusted p-value = 0.005).