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. 2014 May 5;289(24):17020–17029. doi: 10.1074/jbc.M114.563114

FIGURE 5.

FIGURE 5.

Nrf2 is important for ROS-mediated NLRP3 inflammasome activation. A–C, WT, Nrf2−/−, or Nlrp3−/− BMMs were untreated or primed with 400 ng/ml LPS for 3 h followed by the stimulation with rotenone (20 μm) or antimycin (40 μg/ml) for 6 h. IL-1β (A) and IL-6 (B) in supernatants were measured by ELISA. Immunoblotting for caspase-1 and IL-1β were performed in supernatants and cell lysates (C). D, WT BMMs were untreated or pretreated with NAC or DPI for 30 min followed by stimulation with LPS (400 ng/ml) for 6 h. Immunoblotting (left) and densitometric analysis (right) for Nrf2 and NLRP3 inflammasome component molecules were performed. E and F, WT or Nrf2−/− BMMs were untreated or pretreated with NAC or DPI for 30 min followed by stimulation with LPS (400 ng/ml) for 3 h then silica (100 μg/ml) for 6 h (E) or transfected poly(dA:dT) for 4 h (F). IL-1β in supernatants was measured by ELISA. G and H, WT BMMs were treated with LPS for 4 h followed by the stimulation with silica in the absence or presence of NAC or DPI for 4 h. Immunoblotting was performed (G). IL-1β in supernatants was measured by ELISA (H). Values are expressed as the mean ± S.D., and the results are representative of four independent experiments. * p < 0.05, versus controls.