Fig. 7: Mitochondrial ROS is not necessary for CLO97-dependent NLRP3 inflammasome activation.
From: Mitochondrial electron transport chain is necessary for NLRP3 inflammasome activation
a, Intracellular pro-caspase-1 expression in cell lysates from WT and NDI1 BMDMs treated with LPS (100 ng; ml–1) and CL097 (70 μM); N = 4 for all treatments. b, Intracellular caspase-1 (p20 fragment) protein expression in WT and NDI1 BMDMs treated as in a (n = 4 NDI1 all treatments, WT UT, WT LPS, WT LPS + CL097 + antimycin A; n = 3 WT LPS + CL097). c, IL-1β protein levels in cell culture supernatant of WT and NDI1 BMDMs treated with LPS (100 ng ml–1) and CL097 (70 μM). Antimycin (Anti; 100 nM), myxothiazol (Myxo; 100 nM), FCCP (10 μM) or oligomycin (Oligo; 50 nM) were added 30 min before inflammasome activation (n = 5 for all treatments). d, IL-1β protein levels in cell culture supernatant of BMDMs treated with LPS (100 ng ml–1) and ATP (5 mM). MitoTempo (500 μM) was added 30 min before ATP (n = 6 for each condition). e, IL-1β protein levels in cell culture supernatant of BMDMs treated with LPS (100 ng ml–1) and CL097 (70 μM). MitoTempo (500 μM) was added 30 min before the addition of CL097 (n = 6 for each condition). f, IL-1β protein levels in cell culture supernatant of BMDMs treated with LPS (100 ng ml–1) and ATP (5 mM). S1QEL (S1; 1 μM) or S3QEL (S3; 10 μM) was added 30 min before ATP (n = 6 for each condition). g, IL-1β protein levels in cell culture supernatant of BMDMs treated with LPS (100 ng ml–1) and CL097 (70 μM). S1QEL (S1;1 μM) or S3QEL (S3; 10 μM) was added 30 min before CL097 (n = 6 for each condition). h, IL-1β protein levels in cell culture supernatant of BMDMs treated with LPS (100 ng ml–1) and CL097 (70 μM). Paraquat (25 μM) was added 30 min before CL097 (n = 5 WT all treatments; n = 3 NDI1 all treatments). i, Schematic of K+ efflux-dependent (left) and independent (right) NLRP3 inflammasome activation. Error bars represent means ± s.e.m. *P < 0.05, one-way ANOVA with Tukey’s post hoc test for multiple comparisons (b, *P = 0.0454 WT LPS + CL097/NDI1 LPS + CL097, *P = 0.0229 NDI1 LPS + CL097/NDI1 LPS + CL097 + Antimycin A; c, *P < 0.0001 WT LPS + CL097/NDI1 LPS + CL097, *P = 0.0434 NDI1 LPS/CL097/NDI1 LPS + CL097 + Anti, *P = 0.0373 NDI1 LPS + CL097/NDI1 LPS + CL097 + Myxo, *P = 0.0053 NDI1 LPS + CL097/NDI1 LPS + CL097 + FCCP). Parts of this figure were created with BioRender.com.
