Extended Data Fig. 4: Piericidin A decreases NLRP3 inflammasome activation.
From: Mitochondrial electron transport chain is necessary for NLRP3 inflammasome activation
a) Il1b mRNA expression (ΔΔCt) in BMDMs treated with or without LPS (100 ng/mL) for 4 hours, with or without piericidin A (100 nM, 500 nM) (N = 6 for each condition). b) Tnf expression (ΔΔCt) in BMDMs treated as in a (N = 5 for all conditions). c) Il10 mRNA expression (ΔΔCt) in BMDMs treated as in a (N = 5 for all conditions). d) Pro-IL-1β protein expression in cell lysates from BMDMs treated with or without LPS (100 ng/mL), with or without piericidin A (100 nM). (N = 7 LPS; N = 6 UT, LPS + piericidin A; N = 4 LPS + ATP, LPS + piericidin A + ATP). e) Intracellular pro-caspase-1 protein expression in cell lysates from BMDMs treated with or without LPS (100 ng/mL) and ATP (5 mM), with or without piericidin A (100 nM). (N = 8 LPS; N = 7 UT, LPS + piericidin A; N = 6 LPS + ATP, LPS + piericidin A). f) IL-1β protein levels in cell culture supernatant treated with or without LPS (100 ng/mL) and ATP (5 mM), with or without piericidin A (100 nM, 500 nM). (N = 4 for each condition). g) Intracellular caspase-1 (p20 fragment) protein expression in cell lysates from BMDMs treated with or without LPS (100 ng/mL) and ATP (5 mM), with or without piericidin A (100 nM). (N = 6 LPS, LPS + ATP, LPS + piericidin A + ATP; N = 5 UT, LPS + piericidin A.). h) IL-1β protein levels in cell culture supernatant from BMDMs treated with or without LPS (100 ng/mL) and Nigericin (20μM), with or without piericidin A (100 nM, 500 nM). Subsequently, (N = 7 for each condition). i) TNFα protein levels in cell culture supernatant from BMDMs treated as in H. (N = 9 for all conditions). Data are means + /− SEM. * p < 0.05, one-way ANOVA with Tukey test for multiple comparisons (f *p < 0.0001; g *p = 0.0027; h *p < 0.0001).
