FIG. 6.

Ang II-induced alterations of Nox isoform expression in H295R cells. Cells treated with Ang II (10 nM for 6 h) in the absence and presence of AT₁R antagonist (losartan, 10 μM) were analyzed for Nox isoform mRNA expression by real-time PCR (A), and protein expression was analyzed by western blot analysis (B). Cells not treated with Ang II were used as CON. Effects of transfection of siRNA of mock and specific Nox isoforms on corresponding Nox protein expression (C) and superoxide formation in H295R cells (D) are presented. The protein and mRNA data were normalized using β-actin and 18S, respectively. Superoxide was measured in cell homogenates spectrophotometrically by monitoring SOD (2000 U/ml)-inhibitable, NADPH-dependent oxidation of cytochrome C (80 μM). Results derived from six independent experiments are presented as mean±SEM. * and ^#p<0.05; ** and ^##p<0.01. * and ** denote a significant difference between the control and Ang II treated samples. # and ## denote a significant difference between Ang II and Ang II+drug-treated samples. Los, losartan.