Fig. 5.

Aldosterone synthase in Clcn2^op adrenal gland. a–d Quantitative RT-PCR analysis of steroidogenic enzyme mRNA expression in adrenal glands from 12-week-old male and female Clcn2^+/+ (+/+, circles), Clcn2^+/op (+/op, squares), and Clcn2^op/op (op/op, triangles) mice, n ≥ 3 (each point represents one animal). Relative expression levels (compared to Clcn2^+/+ average, reference value of 1 indicated with a dotted line) for a StAR (steroidogenic acute regulatory protein), b Cyp21a1 (steroid 21-hydroxylase), c Cyp11b2 (aldosterone synthase), d Cyp11b1 (11β-hydroxylase). Error bars in a–d, geometric mean ± geometric SD. Statistical analyses was performed on ΔCt values. *p < 0.05, **p < 0.01 (+/op and op/op each compared to +/+ of same sex, male and female analyzed separately, Kruskal–Wallis test, Dunn’s multiple comparison test). e Representative aldosterone synthase/Cyp11b2 staining of paraffin-embedded adrenal gland slices from male and female +/+, +/op, op/op mice at age 12 weeks. f Quantification of Cyp11b2-positive cells observed in e on an automated molecular imaging platform (Vectra, Perkin Elmer). n = 3 animals for each group. Error bars, means ± SEM. *p < 0.05 (+/op and op/op each compared to +/+ of same sex, male and female analyzed separately, one-way ANOVA, Bonferroni multiple comparison test). Scale bar, 100 µM