Fig. 3.

BRET assay between AT₁-R and β-arrestin2 upon agonist stimulation (Stim) in HEK293 cells. HEK293 cells were transfected with AT₁-R–luciferase and β-arrestin2–YFP, and after 24 hours, the cells were exposed to 100 nM AngII (black-filled squares), (A) 10 μM SII-AngII (gray open symbols), (B) 1 μM TRV120023 (TRV3; gray open squares) or 1 μM TRV120027 (TRV7; gray-filled triangles), (C) 10 μM AngIV (gray open triangles), or vehicle (dashed lines) at the indicated time points. (D) HEK293 cells were transfected with the plasmids of the indicated receptor Rluc (WT, wild type, black-filled symbols; DRY/AAY mutant, gray open symbols) and β-arrestin2–YFP, and after 24 hours, the cells were exposed to either 100 nM AngII or vehicle (dashed line) at the indicated time points. The BRET records are the average of at least three independent experiments. Mean values ± S.E.M. are shown (n = 3).