Fig. 5

ER Stress induced NLRP3 inflammasome -mediated pyroptosis activation. The ER stress in HepG2 cells were elicited by chemical ER stressor tunicamycin (TM) for 24 h. a. Cell viability of cells was assessed using the CCK8 assay. b. and c. The mRNA expression of key genes governing ER stress and pyroptosis were detected after 24 h treatment of 0.8 uM TM, and β-ACTIN was used as an internal control. d. Representative western blots of ER stress and pyroptosis makers, and β-ACTIN was used as a protein-loading control. e. HepG2 cells were exposed to TM and plus with TUDCA or OA followed by labeling with anti-GSDMD or anti-NLRP3 antibody and were visualized under a microscope at 400× magnification after 24 h treatment. f. Cells were exposed to TM, or plus with TUDCA, 4-PBA or OA, and cell viability was assessed using the CCK8 assay. g. The mRNA expression of key genes governing ER stress and pyroptosis were detected after 24 h treatment, and β-ACTIN was used as an internal control. h. and i. Representative western blots of GRP78, CHOP, NLRP3 and GSDMD-N after 24 h treatment, and β-ACTIN was used as a protein-loading control. The data are presented as means ± SD for 3–5 biological replicates; *P < 0.05, **P < 0.01, ***P < 0.001vs. BSA;#P < 0.05, ##P < 0.01,vs. TM; ns no significant differences between two connected groups