Fig. 4

Activation of Nrf2 inhibits NLRP3 inflammasome-mediated pyroptosis in BEAS-2B cells. BEAS-2B cells were divided into four groups: control group; TBHQ group; LPS + ATP group; LPS + ATP + TBHQ (20 μM) group. BEAS-2B cells in LPS + ATP + TBHQ group were stimulated with LPS (1 μg/ml) for 4 h and ATP (5 mM) for 30 min following 20 h pretreatment with TBHQ. A, B Hoechst 33,342 (blue)/PI (red) double-fluorescent staining (100 ×) and the rate of PI-positive cell. C, D The protein expression of NLRP3, ASC, CASP1-P20, GSDMD-N were detected by western blot. E LDH release in supernatant. F The mRNA expression of NLRP3, ASC, CASP1, GSDMD were detected by RT-PCR. G, H The production of IL-18 and IL-1β in supernatant were measured by ELISA. I Immunofluorescence staining of GSDMD (original magnification 400 ×). Merged images of DAPI for nucleus (blue) and GSDMD immunofluorescence (green). J The mean fluorescence intensity (MFI) was quantified using ImageJ software. MFI = Integrated Density/Area. Values are expressed as the mean ± SD of three independent experiments. ^*P < 0.05, ^**P < 0.01 compared with the control group, ^#P < 0.05, ^##P < 0.01 compared with LPS + ATP group