FIG. 1.

Ang II-evoked changes in aldosterone production and CYP11B2 activity, mRNA, and protein expression in human H295R cells. (A) Aldosterone biosynthetic pathway. Cells grown to 90% confluence were treated with indicated concentrations of Ang II for 6 h. Cells not treated with Ang II served as control (CON). Changes in aldosterone levels in the culture medium (B) and CYP11B2 activity in cell homogenates (C) as a function of concentration of Ang II were determined using procedures as described in Materials and Methods section. Ang II (10 nM)-induced changes in CYP11B2 activity as a function of incubation time (D), CYP11B2 protein as determined by western blot (E), and CYP11B2 mRNA expression as assessed by real-time PCR (F) are shown. The protein and mRNA data were obtained with cells treated with either vehicle or Ang II for 6 h and normalized using β-actin and 18S, respectively. Results derived from six independent experiments are presented as mean±SEM. *p<0.05, **p<0.01, ***p<0.001 and n.s., not significant. Ang II, angiotensin II; CYP11B2, aldosterone synthase; PCR, polymerase chain reaction.