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. 2004 Dec;24(24):10703–10717. doi: 10.1128/MCB.24.24.10703-10717.2004

FIG. 4.

FIG. 4.

Silencing of Nox-4 expression by RNA interference in SMCs. (A) Cells were transfected with scrambled siRNA or Nox-4 siRNA duplexes. After incubation for 24 h in complete medium, SMCs were treated with 0 or 40 μg of 7-Kchol/ml for 16 h. Nox mRNA expression levels were then determined by real-time RT-PCR. Bars, indicating the relative expression of Nox versus α-actin mRNA levels, are means ± the SD from three independent experiments. ∗, P < 0.01. (B) SMCs transfected with scrambled or Nox-4 siRNAs were treated with 0 or 40 μg of 7-Kchol/ml for 16 h. The expression of Nox-4 was detected by Western blot analysis with the anti-Nox-4 antibody. The bars represent the mean densitometric value of Nox-4 over α-actin from three separate experiments for each of the experimental conditions tested. (C) ROS production (chemiluminescence assay) was measured on SMCs transfected with scrambled or Nox-4 siRNAs incubated with 0 or 40 μg of 7-Kchol/ml for 16 h. A combination of PEG-SOD (10 μg/ml) and PEG-catalase (10 U/ml) was added 5 min before the chemiluminescence assay. The chemiluminescence intensity values reported are means ± the SD from three separate experiments performed in triplicate. ∗, P < 0.01.