Fig. 8: CHOP upregulated GSDME expression, while its knockdown ameliorated pyroptosis, mitochondrial damage, and inflammation in renal tubular epithelial cells.

A Western blot analysis showed protein expression of GSDME, GSDME-N, Cleaved-Caspase3 in different groups. B JC-1 staining representation of mitochondrial disability in different groups, n = 3/group, Scale bar: 100 μm. C, D DCFH staining of ROS and C11 staining of lipid peroxidation levels, which confirmed the release of ROS, n = 3/group, Scale bar: 100 μm. E Representative co-localization images of the mitochondrial marker MitoTracker (labeled in red) and Cytochrome c(cyt-c) (labeled in green) in HK-2 cells; the protein fluorescence intensity/area in each group are shown, n = 3/group, Scale bar: 100 μm; Scale bar: 20 μm (magnification). F Real-time PCR detected mRNA levels of IL-1β, IL-6, TNF-a, MCP-1 in different groups. G Western blot analysis showed protein expression of P65, PP65 in different groups. H Chromatin immunoprecipitation analysis with antibodies against CHOP or IgG, soluble chromatin from PMs, and primers targeting the region spanning the binding site in the GSDME promoter in different groups, n = 3/group. For all panels, p value was determined by unpaired two-tailed Student’s t test or one-way ANOVA with Bonferroni post hoc test for multiple comparisons. Data are expressed as mean ± SEM. Quantification on the blots derive from samples of the same experiment and gels/blots were processed in parallel. ns not significant p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.