FIGURE 3.

Manoalide has no effects on potassium efflux, chloride efflux or mitochondrial damage and inhibits the NLRP3 inflammasome activation independently of PLA2/PLC signaling. (A–D) PLA₂ and PLC expression were knock down by siRNA in BMDMs. Then, the BMDMs were primed with LPS for 3 h and then were pretreated with 500 nM manoalide before stimulation with nigericin for 15 min (A) Pla2g4a, (B) Pla2g7, (C) Pla2g15 and (D) Plcg2 mRNA in BMDMs transfected with siRNA or control siRNA and ELISA of mature IL-1β in supernatant. Qualification of potassium efflux (E) and chloride efflux (F) in BMDMs primed with LPS and then were pretreated with 125, 250, 500 nM manoalide before stimulation with nigericin. (G) Confocal microscopy analysis of LPS-primed BMDM pretreated with 500 nM manoalide before stimulation with nigericin, then stained with MitoSOX red, Mitotracker red and DAPI. Data are derived from three independent experiments (A–F) and displayed as mean and SEM (n = 3) or represent three independent experiments (G). Statistically significant difference was calculated by unpaired Student’s t-test: NS, not significant.