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. 2015 Apr 2;156(6):2138–2149. doi: 10.1210/en.2014-1866

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Figure 8. — Knockdown of calpain-10 inhibited AngII-induced CYP11B2 mRNA expression levels and aldosterone production in nucleofected human adrenocortical carcinoma (HAC15) cells. HAC15 cells were transfected by nucleofection with 40nM pooled calpain-10 siRNA or 40nM control scrambled siRNA with GFP and incubated for 48 hours. Before experiments, medium was replaced with low-serum medium overnight followed by treatment in the presence and absence of 10nM AngII for 24 hours. Aldosterone levels were assayed by a solid-phase RIA (A), RNA was isolated and reverse transcribed for qRT-PCR assay of calpain-10 (B) and CYP11B2 (C) expression. The data are expressed as the fold over the scrambled-siRNA control (sc-sirna) and represent the mean ± SEM of at least 3 separate experiments performed in duplicate; ***, P < .001; *, P < .05 vs scrambled siRNA; ƒƒƒ, P < .001; ƒƒ, P < .01; ƒ, P < .05 vs AngII plus scrambled siRNA by ANOVA followed by a Student-Newman-Keuls post hoc test.