Fig. 3

Functional characterization of AT₁R-AT₂R heteromer in HEK-293T cells. HEK-293T cells were transfected with cDNAs for AT₁R (1 μg) and/or AT₂R (1 μg). Cells were pretreated (15 min) with selective receptor antagonists (300 nM candesartan for AT₁R or 1 μM PD123319 for AT₂R receptors) and subsequently treated with selective agonists (100 nM angiotensin II for AT₁R and 300 nM CGP-42112A for AT₂R receptors). a–c ERK1/2 phosphorylation was analyzed using an AlphaScreen®SureFire® kit (Perkin Elmer). Values are the mean ± S.E.M. of 5 independent experiments performed in duplicates. One-way ANOVA followed by Bonferroni’s multiple comparison post-hoc test were used for statistical analysis. Signaling output was the dependent variable and the different treatments were the independent variables (*p < 0.05, **p < 0.01, ***p < 0.001; versus vehicle treatment (basal)). d–f DMR tracings represent the picometer-shifts of reflected light wavelength over time. Values are the mean ± S.E.M. 8 independent experiments performed in triplicates