Figure 1.

AngII-mediated internalization and trafficking profiles of WT and mutant AT1R with β-arrestin. A and B, HEK293 cells were transfected with RlucII-tagged WT or mutant AT1R and PM-anchored rGFP-CAAX (A) or endosomally anchored rGFP-FYVE (B). C, HEK293 cells were transfected with WT or mutant AT1R, β-arrestin2-RlucII, and rGFP-FYVE. Cells were stimulated with 1 μm AngII for up to 1 h in time course experiments (left panels) or stimulated with indicated AngII concentrations for 1 h in concentration-response experiments (right panels). For A, the percentage change in basal BRET is reported, whereas for B and C, the BRET change was normalized and reported as percentage of WT maximal response. Data are represented as mean ± S.D. of triplicates from three to six independent experiments, and for left panels, two-way ANOVA followed by Dunnett's multiple comparisons tests were performed for the last time point where *** = p < 0.001, ** = p < 0.01, * = p < 0.05, n = 3–6.