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. 2016 Jan 20;291(11):5688–5707. doi: 10.1074/jbc.M115.669952

FIGURE 5.

FIGURE 5.

Delineation of the source of ROS generation in the MIOX-overexpressing cell line and effect of NOX siRNA on ROS under high glucose ambience. Mito Q and DPI, both known inhibitors of ROS generated from mitochondria and NADPH oxidase, respectively, were used. Both inhibited DCF-DA-associated fluorescence, as delineated by flow cytometric analyses (A–E), suggesting that the ROS are derived from mitochondria as well as the NADPH oxidase system. Next, we addressed the question of whether NOX4-specific inhibition could reduce the generation of ROS in MIOX-overexpressing cells under high glucose ambience. NOX4 siRNA inhibited generation of ROS, both under basal conditions and under high glucose ambience (F–L). However, siRNA treatment could not reduce the mean fluorescence intensity to basal levels, suggesting an additional source of ROS generation (e.g. mitochondria) in MIOX-overexpressing cells under high glucose ambience (n = 4; *, p < 0.01; #, p < 0.05). Error bars, S.D.