Fig. 2

ACSS2 contributes to the inflammatory response and pyroptotic cell death in LPS-treated HK-2 cells. A Representative immunohistochemical staining and Western blotting analysis of ACSS2 in the kidneys of control and LPS-induced AKI mice. (Scale bars, 50 μm; n = 4). B Immunofluorescent staining of ACSS2 in HK-2 cells with or without LPS treatment (green, ACSS2; blue, DAPI; scale bars, 200 μm). C Western blotting analysis of ACSS2 protein in HK-2 cells treated with LPS for 24 or 48 hours (n = 3). D Relative mRNA expression levels of the pro-inflammatory cytokines (MCP-1, IL-6, and IL-1β) in HK-2 cells with or without pre-treatment with an ACSS2 inhibitor (ACSS2i) prior to LPS treatments for 24 or 48 hours (n = 3). E Relative mRNA expression level of kidney injury molecule-1 (KIM-1) in HK-2 cells with or without pre-treatment with ACSS2i prior to LPS treatments for 24 or 48 hours (n = 3). F The mRNA and protein levels of ACSS2 in HK-2 cells transfected with an ACSS2-specific siRNA or scrambled siRNA prior to LPS treatments for 24 hours (n = 3). G Relative mRNA expression levels of the pro-inflammatory cytokines (MCP-1, IL-6, and IL-1β) in HK-2 cells transfected with an ACSS2-specific siRNA or scrambled siRNA prior to LPS treatments for 24 hours (n = 3). H: TEM observation of the ultrastructure of HK-2 cells with or without pre-treatment with ACSS2i prior to LPS treatments for 24 hours (Scale bars, 5 μm). I: Immunofluorescence detection of the expression of cleaved GSDMD in HK-2 cells with or without LPS (1 μg/mL) and with or without co-treatment with ACSS2i (10 μmol/L) for 24 hours (Scale bars, 100 μm). J: Western blotting analysis of GSDMD 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 (n = 3). Data were presented as mean ± SD. ^*P < 0.05, ^**P < 0.01, ^***P < 0.005 vs. Ctrl; ^#P < 0.05, ^##P < 0.01, ^###P < 0.005 vs. LPS or LPS + siControl; ^$P < 0.05 vs. LPS for 48 hours