FIG. 6.

Aldosterone-induced activation of Nrf2 by oxidants in LLC-PK1 cells depends on the MR, protein kinase C, NADPH oxidase, nitric oxide synthase, and intracellular calcium. (A) The involvement of the MR was evaluated by measuring Nrf2-DNA binding (EMSA) in nuclear fractions from aldosterone-treated cells that were simultaneously incubated without or with either 500 nM eplerenone (Ep) or 500 nM mifepristone (Mi). LLC-PK1 cells were incubated in the absence (C) and the presence of 100 nM aldosterone (Ald) for 4 h. Aldosterone-treated cells were simultaneously incubated with (B) 1 μM DPI (DPI), 50 μM apocynin (Apo), 1 μM VAS2870 (VAS) and 1 μM Ro320432 (Ro), (C) 50 μM L-NAME (LN), 10 μM W-7 (W7), or (D) 10 μM bapta-AM (Bapta). (A–D) Shown are mean values±SEM, *p≤0.05 versus the control group, ^#p≤0.05 versus the aldosterone-treated group, tested by ANOVA with subsequent post-hoc comparisons by Scheffé. DPI, diphenyleneiodonium chloride; W-7, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride; VAS2870, 3-benzyl-7-(2-benzoxazolyl)thio-1,2,3-triazolo[4,5-d]pyrimidine).