Fig. 6.
Effect of Cav3.2196Leu and Cav3.21549Ile on mRNA expression of genes involved in aldosterone biosynthesis. 24 h post transfection, cells were serum deprived (0.1% Ultroser G) for 24 h and then incubated for another 24 h with fresh serum deprived medium in absence (basal) or presence of 10 nM of AngII (A-II 10 nM) or 12 mM K+ (K+ 12 mM). Cells were collected for RNA extraction, and real time RT-qPCR was performed. mRNA levels of StAR, HSD3B1, HSD3B2, CYP21A2 and CYP11B2 were normalized for CACNA1H expression to correct for transfection efficiency. Normalization for RNA quantity, and reverse transcriptase efficiency was performed against three reference genes (geometric mean of the expression of 18S RNA, HPRT, and GAPDH). Data are represented as fold increase vs WT in basal condition. i) **P < 0.01 or *P < 0.05 vs Cav3.2WT basal; ii) ***P < 0.001 or **P < 0.01 or *P < 0.05 vs Cav3.2WT K+, n = 9–12.