Fig. 6

Resatorvid mitigates inflammatory responses of macrophages via NLRP3/ROS signaling. A Immunofluorescence and quantification of the expression of NLRP3 in Raw264.7 macrophages treated as indicated for 24 h. M1 macrophages were induced by 100 ng/ml LPS and 20 ng/ml INF-γ, one-way ANOVA, mean ± SEM, n = 3 (###p < 0.001 vs M0 group; *p < 0.05, ***p < 0.001 vs M1 group). B, C The production of ROS in Raw264.7 macrophages as specified was labeled by cell-permeable dihydroethidium (DHE) fluorogenic probes (B) and quantified by the average optical density of red fluorescence (C), one-way ANOVA, mean ± SEM, n = 3 (##p < 0.01 vs M0 group; *p < 0.05, **p < 0.01 vs M1 group). ELISA analysis detecting the concentration of cytokines (IL-1β, TNF-α, and IL-6) in the supernatant of Raw264.7 macrophages treated as indicated for 24 h, one-way ANOVA, mean ± SEM, n = 3 (##p < 0.01, ###p < 0.001 vs M0 group; *p < 0.05, **p < 0.01 vs M1 group, ns indicates no significant difference). E, F Western blotting analysis (E) and quantification (F) of the production of NLRP3 and proinflammatory factors (iNOS, NF-κB, COX-2, IL-1β, IL-6) in lysates of Raw264.7 macrophages treated as indicated, Student’s t test, mean ± SEM, n = 3 (#p < 0.05, ##p < 0.01 vs M0 group; *p < 0.05, **p < 0.01 vs M1 group, ns indicates no significant difference)