Figure 4.

NAMPT peptides inhibit the signal 1 and 2 activation of the NLRP3 inflammasome. (A–C) BMDMs were stimulated with LPS for 4 h, pretreated with rNAMPT for 1 h (A,B) or 18 h (C). (A) WCLs were used for IB with αIkB-α, αP-IkB-α or αActin. (B) FACS analysis for H₂O₂ (probe for 2′, 7′-dichlorofluorescin diacetate) or O₂- (probe for Dihydroethidium). Quantitative analysis of mean fluorescence intensities of H₂O₂ and O₂- (upper). NADPH oxidase activity (lower). (C) Culture supernatants were harvested and analyzed for cytokine ELISA. (D,E) LPS-primed BMDMs were treated with rNAMPT for 1 h, and then activated with ATP for 30 min. (D) ELISA for IL-1β and IL-18. (E) IB in SN with αIL-1β p17, αIL-18 p18 or αCasp1 p10 and WCL with αPro-IL-1β, αPro-IL-18, αPro-Casp1, or αActin. The data come from seven independent experiments that yielded comparable results (A,E). The data come from three independent experiments that yielded comparable results. (B–D). Data are given as means ± SD of three experiments. Significant differences (* p < 0.05; ** p < 0.01; *** p < 0.001) versus LPS + PBS.