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. 2020 Apr 27;11(4):295. doi: 10.1038/s41419-020-2459-3

Fig. 4. Activation of the caspase-9/caspase-3 pathway triggers the GSDME-mediated pyroptosis in response to CAP treatment.

Fig. 4

a–d The CAP-induced pyroptosis was repressed in PC9 and SGC-7901 cells pre-treated with pan-caspase inhibitor zVAD (30 µM) for 2 h following 40 s CAP exposures. a Representative microscopic images in which red arrowheads indicated large bubbles emerging from the plasma membrane. Scale bar, 25 µm. b Apoptosis- and pyroptosis-related proteins including PARP, cleaved-PARP, GSDME, GSDME-N and pro-CASP-3 detected by western blotting. c Release of LDH in the culture supernatant. d Cell death assessed by measuring annexin V-FITC- and PI-stained cells. e Apoptosis and pyroptosis-related proteins as indicated were detected after CAP treatment by western blotting in PC9 and SGC-7901cells. f, g Knocking down of caspase-3 (CASP-3) or caspase-9 (CASP-9) reduced the occurrence of apoptosis and pyroptosis induced by CAP exposure. Apoptosis and pyroptosis-related proteins as indicated were detected at 24 h after CAP exposures for 40 s in PC9 cells transfected with caspase-3 siRNA (f) and caspase-9 siRNA (g), respectively. h Apoptosis and pyroptosis-related proteins as indicated were detected at 24 h after CAP exposures for 40 s in PC9 cells pretreated with caspase-9-specific inhibitor zLEHD (30 µM). All the data are presented as the mean ± SD from three independent experiments. *p < 0.05, **p < 0.01.