Fig. 5
Granzyme B cleaves GSDMC in response to DdBIC stimulation. Melanoma A375 cells were treated with DdBIC (20 μM) for 6 h to determine the level and activity of granzyme B (GZMB), for 7 h to determine the interaction between GZMB and GSDMC, or for 8 h to assess pyroptotic features, unless otherwise specified. a Cells were cotreated with 3,4-DCI (10 μM) for 8 h to detect pyroptosis. b GZMB was transfected into cells, and the GZMB level in the cytosolic fraction was detected in response to DdBIC stimulation. c GZMB and GSDMC were overexpressed for the TurboID-based assessment of the interaction between GZMB and GSDMC. d GZMB was knocked down to assay pyroptosis. e GZMB and GZMBS183A were transfected into GZMB-knockdown cells to detect pyroptosis. f The cleavage of GSDMC by recombinant GZMB was detected in vitro. g, h Cells were cotreated with ISRIB and DdBIC or with PERK knockdown to determine the mRNA (g) or protein (h) levels of GZMB and GZMB activity. i GSDMC and GSDMCD276A were reintroduced into GSDMC-knockdown cells to detect pyroptosis. Statistics: two-way ANOVA with Tukey’s test to a, d, g and h; one-way ANOVA with Tukey’s test to e and i. P values are shown
