Fig. 2

Downregulation of GSDMD alleviates alcohol-induced hepatocyte pyroptosis. GSDMD downregulation shRNA (sh-GSDMD) or negative control shRNA (sh-NC) were transfected into hepatocytes. After transfection, hepatocytes were stimulated with or without 40% alcohol for 48 h. A The gene expression of GSDMD in hepatocytes was determined by qPCR. B The protein expression of GSDMD-N in hepatocytes was determined by western blot. C The levels of IL-1β and IL-18 in the culture supernatants of hepatocytes were measured by ELISA kits. D The expressions of NLRP3, ASC, cleaved Caspase 1 and pro-IL-1β in hepatocytes were determined by western blot. E The LDH level in the supernatants of hepatocytes was measured. F Fluorescence microscopy analysis of propidium iodide (PI) internalization expressed as PI positive cells percentages (%). Data are presented as mean ± SD. *p < 0.05, **p < 0.01