Fig. 5

DS negatively regulated the formation of NLRP3 inflammasome complex through mitophagy in BV-2 microglia. BV-2 microglia were first pre-stimulated by LPS (100 ng/ml) for 3.5 h, then treated for 1h with indicated concentrations of DS and 3-MA (5 mM) for 30 min or not followed by stimulation with nigericin (5 μM) for 6 h, 12 h, and 24 h, which was labeled with a triangle at nigericin (5 μM) treatment. a Immunoblot analyses of caspase-1 p10, IL-1β p17 in culture supernatants (SN) and NLRP3, pro-caspase-1, pro-IL-1β in lysates (LYS) were shown. b Immunofluorescence staining show the expression of NLRP3 (green) and ASC (Red) in BV-2 microglia after stimulation with nigericin for 12 h. c, d Caspase-1 activity was detected by flow cytometry in BV-2 microglia treated with DS for 12 h using caspase-1/PI double-labeled staining method. Data are expressed as means ± SEM (n = 3 in each group). Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc analysis where ^###P < 0.001 vs. control group; ^***P < 0.001 vs. LPS + Nigericin group; ^$$$P < 0.001 vs. LPS + Nigericin + 30 μM DS group