Fig. 3.

Caspase-1-induced apoptosis depends on caspase-3. a–c Gsdmd-KO and Gsdmd/Casp3-DKO CL26-iCasp1 cells were treated with AP20187 for the indicated times. Western blot detection of caspase-3 and GSDME in cell lysates (a). LDH release from the AP20187-treated cells (b). PI uptake and PS exposure analyzed by flow cytometry (c). Percentages of Annexin V⁺ cells are shown in Supplementary Fig. 4b. d, e Gsdmd-KO and Gsdmd/Casp3-DKO RAW264.7 cells were infected with S. Typhimurium as in Fig. 1f (d) or Fig. 1e (e). Caspase-3 in cell lysates detected by Western blotting (d). Microscopic images (e). f, g Gsdmd-KO and Gsdmd/Gsdme-DKO CL26-iCasp1 cells were treated with AP20187, and cell death was monitored by LDH release (f) and flow cytometry (g). Percentages of Annexin V⁺ PI⁺ cells and Annexin V⁺ PI⁻ cells are shown in Supplementary Fig. 5d. In b and f, graphs depict the mean ± SD of triplicate cultures, and individual data values are plotted (b, f). Statistical significance was determined using an unpaired Student’s t-test (f). ***p < 0.001. Data are from one representative of three independent experiments with similar results (a–g). Source data are provided as a Source Data file. (See also Supplementary Figs. 4 and 5.)