FIGURE 5. NADPH oxidase-derived ROS do not contribute to cyclic stretch-induced NLRP3 inflammasome activation.
A. AMs isolated from gp91phox−/−mice show no change in IL-1β release following stretch. Wild type and gp91phox−/−AMs were subjected to 20% cyclic stretch for 4 h. IL-1β release in the culture supernatants (SN) of AMs and levels of Pro-IL-1β, Pro-Casp-1 and Casp-1 in cell lysate (Lys) were determined by Western blot analysis. B. The levels of IL-1β in cell-culture media were measured by ELISA (n=3). C. Representative histograms of flow cytometry experiments demonstrating the effects of cyclic stretch on mitochondrial ROS generation in gp91phox−/− and wild type AMs. D. Quantitative data showing changes in mean fluorescent intensity (MFI) of MitoSOX following stretch (n=3). *p < 0.05 vs. the control (static) group.