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Figure 4. Endogenous ROS induced by NADPH oxidase is specifically involved in LPS signaling and proinflammatory responses in Prx IIdeficient cells. (A) After preincubation for 30 min with 10 and 30 mM NAC, 10 and 20 µM DPI, 10 and 100 µM allopurinol, 1 and 10 µM rotenone, 100 and 500 µM L-NMMA, or 100 and 500 µM L-NAME, BMDMs were stimulated with 1 µg/ml LPS for 30 min. The cells were harvested and subjected to Western blot analysis for I B , phosphorylated ERK 1/2, and p38 MAPK. The same blots were washed and blotted for ß-actin as the loading control. Data shown are representative of three independent experiments that gave similar results. (B) The experimental conditions followed the pattern outlined in A. Culture supernatants were harvested after stimulation with LPS for 18 h, and the TNF- and IL-6 expression levels were measured by ELISA. Data shown are the mean ± SD of three experiments. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 compared with WT cell cultures stimulated with LPS. MC, media control.
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