Figure 7.

Calpain-10 overexpression increased AngII-induced aldosterone production but had no effect on AngII-induced CYP11B2 mRNA expression levels in human adrenocortical (HAC15) cells. Cultured HAC15 cells were incubated for 6 hours with adenovirus expressing GFP or recombinant calpain-10. After 6 hours of infection, low-serum medium was replaced with fresh medium and the cells incubated for an additional 18 hours. Cells were then treated with or without 10nM AngII for 24 hours. Aldosterone levels were assayed by a solid-phase RIA (A) and cells processed for Western blotting using calpain-10 antibody (B). RNA was isolated and reverse transcribed for qRT-PCR assay of the expression of calpain-10 (C) and CYP11B2 (D). All data are expressed as fold over the vector-infected control and represent the mean ± SEM of at least 3 separate experiments performed in duplicate; ***, P < .001; **, P < .01; *, P < .05 vs the vector (GFP-infected); ƒƒƒ, P < .001; ƒƒ, P < .01, vs AngII + vector by ANOVA followed by a Student-Newman-Keuls post hoc test.