Mammalian Target of Rapamycin Regulates Nox4-Mediated Podocyte Depletion in Diabetic Renal Injury

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FIG. 3.
FIG. 3.

mTOR regulates HG-induced ROS production, Nox4 mRNA and protein levels, and NADPH oxidase activity. Mouse podocytes were serum deprived overnight and pretreated with rapamycin (Rapa) (10 nmol/L) for 1 h before incubation with HG for 48 h. A: ROS generation measured by DCF with a multiwell fluorescence plate reader. B: Relative mRNA levels of Nox4 in control and treated podocytes. C: Representative Western blots of Nox4 and β-actin levels. D: Histograms showing quantitation of Nox4/β-actin from four different experiments. E: NADPH-dependent superoxide generation. In parallel experiments, mouse podocytes incubated in NG or HG were transfected with nontargeting siRNA (Scr) or with SMARTpool of siRNA targeting mTOR (simTOR). F: ROS generation measured by DCF with a multiwell fluorescence plate reader. G: Relative mRNA levels of Nox4 in control and treated podocytes. H: Representative Western blot of Nox4 and β-actin levels. I: Histograms showing quantitation of Nox4/β-actin from four different experiments. J: NADPH-dependent superoxide generation. All values are the mean ± SE from four independent experiments. *P < 0.05 vs. control; #P < 0.05 vs. HG.

This Article

  1. Diabetes vol. 62 no. 8 2935-2947