Fig. 5: Activated Gasdermin E promoted pyroptosis and localized to mitochondria in renal tubular epithelial cells, which enhanced caspase3 activation and formed a positive feed-forward damage response loop in renal tubular epithelial cells. | Cell Death & Disease

Fig. 5: Activated Gasdermin E promoted pyroptosis and localized to mitochondria in renal tubular epithelial cells, which enhanced caspase3 activation and formed a positive feed-forward damage response loop in renal tubular epithelial cells.

From: CHOP-mediated Gasdermin E expression promotes pyroptosis, inflammation, and mitochondrial damage in renal ischemia-reperfusion injury

Fig. 5

A Representative light microscope images of cells. Red arrows indicate large bubbles from the plasma membrane. White and black arrows represent cells in the normal and model groups, respectively. n = 3/group, scale bar: 20 μm. B Representative transmission electron microscopy (magnification ×10,000; scale bar = 2 μm) of cells between control and HR. Red arrowheads indicate large bubbles and pores emerging from the plasma membrane. Blue arrowhead indicate spillover organelles. C Representative cell viability and LDH release (OD values), respectively. D Western blot analysis showed protein expression of KIM1, GSDME, GSDME-N, Cleaved-Caspase3. E Real-time PCR detected mRNA levels of IL-1β, IL-6, TNF-a, MCP-1. F Western blot analysis showed protein expression of PP65, P65. G Representative co-localization images of Dil (labeled in red) and GSDME (labeled in green) in HK-2 cells; Scale bar: 100 μm; Scale bar: 20 μm (magnification). H Representative co-localization images of the mitochondrial marker MitoTracker (labeled in red) and GSDME (labeled in green) in HK-2 cells; White arrows indicate mitochondrial and GSMDE co-localization sites. Fluorescence co-localization display showed correlation analysis, n = 3/group, Scale bar: 100 μm; Scale bar: 20 μm (magnification). I JC-1 staining representation of mitochondrial disability, n = 3/group, Scale bar: 100 μm. J DCFH staining of ROS level, n = 3/group, Scale bar: 100 μm. K 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). For all panels, p value was determined by unpaired two-tailed Student’s t test. 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.

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