Fig. 3

Promotion of NLRP3 inflammasome activation and GSDMD cleavage by ACSS2 depends on NF-κB signaling activation. A The immunofluorescence staining of NF-κB p65 was detected in HK-2 cells which were treated with LPS (1 μg/mL) or vehicle along with ACSS2i (10 μmol/L) for 24 hours. (green, p65; blue, DAPI; scale bars, 100 μm). B The effects of ACSS2 siRNA (siACSS2) on the protein expression and phosphorylation of p65 in HK-2 cells stimulated with LPS (1 μg/mL) were analyzed by Western blotting (n = 3). C and D Western blotting and immunofluorescence analyses of NLRP3 in HK-2 cells which were stimulated with LPS (1 μg/mL) or vehicle and co-incubated with ACSS2i (10 μmol/L) for 24 or 48 hours (red, NLRP3; blue, DAPI, scale bars, 100 μm; n = 3). E and F Western blotting analyses of the protein expression of (E) NLRP3 or (F) pro-caspase-1, caspase-1, pro-IL-1β, and IL-1β in HK-2 cells which were treated with LPS (1 μg/mL) or vehicle and ACSS2 siRNA (siACSS2) for 24 hours (n = 3). G The expression of IL-1β in mouse kidney was determined by immunofluorescence (green, IL-1β; blue, DAPI; scale bars, 50 μm). Data were presented as mean ± SD. ^*P < 0.05, ^***P < 0.005 vs. control; ^#P < 0.05, ^##P < 0.01 vs. LPS or LPS + siControl