Fig. 6.

Bid plays a critical role in caspase-1-induced apoptosis. a–d WT, Gsdmd-KO or Gsdmd/Bid-DKO CL26-iCasp1 cells were treated with AP20187 for the indicated times. Cytochrome c, Bid, GAPDH (cytosolic marker) and MTCO1 (mitochondrial marker) in the whole cell lysates or cytosol fractions were detected by western blotting (a). Bid in the cell lysates and culture supernatants (sup) was detected by Western blotting (b). LDH release into culture supernatants (c). Caspase-3/7 activity in the cell lysates. e–g Gsdmd-KO CL26-iCasp1 cells and Gsdmd/Bid-DKO CL26-iCasp1 cells stably transfected with GFP, GFP-Bid, or GFP-Bid mutants were treated with AP20187 for the indicated times. Cleaved caspase-3 and Bid in cell lyesates were detected by western blotting (e). LDH release (f). Flow cytometric analysis of PI uptake and PS exposure (g). Percentages of Annexin V⁺ cells are shown in Supplementary Fig. 9d. In c, d, and f, graphs depict the mean ± SD of triplicate cultures, and individual data values are plotted. Statistical significance was determined using the unpaired Bonferroni’s multiple comparisons test. n.s., not significant; ***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. 8 and 9.)