Downregulation of TRPC6 expression by sildenafil leads to reduced TRPC6–mediated Ca2+ influx into the podocyte. Downregulation of TRPC6 expression by sildenafil.

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Downregulation of TRPC6 expression by sildenafil leads to reduced TRPC6–mediated Ca2+ influx into the podocyte. Downregulation of TRPC6 expression by sildenafil leads to reduced TRPC6–mediated Ca2+ influx into the podocyte. Intracellular Ca2+ concentration was determined by fura-2 ratiometry. Cultured podocytes transfected with a scrambled siRNA construct were pretreated with sildenafil or 8-Br-cGMP with or without the PKG-1 antagonist KT5823 for 24 hours. (A) After removal of the specific media, cells were exposed to 100 μM 1-oleoyl-2-acetyl-sn-glycerolin (OAG) to activate the TRPC6 ion channel. (B) The same experimental setup was tested in TRPC6 KD podocytes. The arrows indicate the time points of OAG application; n=5 in two independent experiments. Statistical significance was determined using a repeated measurement test. *P<0.05 versus vehicle-treated cells. Ramon Sonneveld et al. JASN 2017;28:1491-1505 ©2017 by American Society of Nephrology