Figure 4.

Evaluation of the contribution of G-protein-dependent and independent mechanisms in the AT₁-receptor stimulation on induced changes in DUSP levels in vascular smooth muscle cells. Serum-depleted VSMCs were incubated with 1 µM YM-254890 (YM) or DMSO as a control for 30 min, then the cells were exposed to either 100 nM AngII (red columns) or 50 ng/mL EGF (blue columns) or vehicle (white columns) for 2 h (A–C). The serum-starved VSMCs were exposed to either the vehicle (white columns) or 100 nM AngII I (red columns) or 3 µM TRV120023 (beige columns) for 2 h (D–F). RNA was isolated from VSMCs, then converted to cDNA. cDNA levels of DUSP5 (A,D), DUSP6 (B,E) and DUSP10 (C,F) were measured by qRT-PCR. Standardization was established against the GAPDH housekeeping gene. Mean values ± SE are shown. Significance was determined via multiple linear regressions. p < 0.05 was considered as statistically significant. *: statistically significant from vehicle stimulation. #: statistically significant from DMSO pretreated agonist-induced response (A–C). In the case of D–F, significance was determined with a one-way ANOVA-test (* p < 0.05). The values are from four or five independent experiments (n = 4–5).