Fig. 8: MafB upregulation diminishes the NLRP3 inflammasome activation.

a, b PMs were pre-treated with vehicle or the RXR activator LG100268 (LG268) (1 µM) for 1 day, followed by LPS treatment for 3 h. The cells were then treated with or without ATP (5 mM) or Nigericin (5 µM), or transfected with poly(dA:dT) or Flagellin. Levels of the indicated protein in supernatants and cell extracts was determined by western blotting (a). Levels of supernatant TNF-α and IL-6 were determined by ELISA (b). Vertically stacked bands originate either from a single membrane or from a replicate membrane, with the same sample loadings. c MafB+/+ and MafB−/− PMs were pre-treated with LG268, followed by LPS priming and ATP activation. ELISA for IL-1β was performed. d MafB+/+ PMs were treated with various concentrations of LG268 for 1 day. Western blotting was performed for the indicated proteins. e Levels of the indicated proteins in control siRNAs or MafB siRNAs transfected PMs treated with or without LG268 for 1 day. f MafB+/+ and MafB−/− PMs were treated with or without LG268 for 1 day. g PMs were pre-treated with vehicle or the RXR activator LG268 for 1 day, followed by LPS treatment for 4 h. The cells were then stimulated with ATP for 30 min, and stained with 100 nM MitoTracker Deep Red and 100 nM MitoTracker Green for additional 20 min. Cells were collected, and flow cytometric analysis was performed to determine the mitochondria damage. h Primed PMs were incubated with MitoSOX for 15 min, followed by ATP stimulation. mtROS levels were determined. Mean ± SD; **P < 0.01, ***P < 0.001. Representative of three independent experiments.