Fig. 6

ACSS2 induces KLF5-mediated p65 activation and downstream cell pyroptosis in LPS-induced kidney tubular injury. A RNA-seq analysis of kidney tissues from ACSS2 gene knockout (KO) mice and wild-type mice was performed. The colored dots indicated differentially expressed genes (DEGs). B Relative protein expression levels of KLF5 in the mouse kidney (n = 3). C Representative immunofluorescent images of KLF5 and LTL in the renal cortex of mice (green, LTL; red, KLF5; blue, DAPI; Scale bars, 100 μm). D Western blotting was used to compare expression of KLF5 in HK-2 cells treated with LPS (1 μg/mL) or vehicle and with ACSS2 siRNA or scrambled siRNA for 24 hours (n = 3). E Immunofluorescence staining was used to detect KLF5 (green, KLF5; blue, DAPI; scale bars, 50 μm). F and G HK-2 cells were pre-treated with ML264 (10 μmol/L) or vehicle for 1 hour, then with LPS (1 μg/ml) or vehicle for 24 hours. Western blotting was used to analyze (F) the expression of KLF5 or (G) the phosphorylation of p65, the expression of NLRP3, and the cleavage of GSDMD (n = 3). H The levels of cleaved GSDMD were determined by immunofluorescence staining (green, cleaved GSDMD; blue, DAPI; scale bars, 50 μm). Data were presented as mean ± SD. ^*P < 0.05, ^***P < 0.005 vs. Ctrl, siControl or LPS + vehicle; ^#P < 0.05, ^##P < 0.01 vs. LPS + siControl or LPS + vehicle