Extended Data Fig. 10: Defective palmitoylation or depalmitoylation sensitizes mice to bacterial infection in a GSDMD-dependent manner.

a, Eight-week-old male mice were injected intraperitoneally with LPS (54 mg/kg body weight) for 3 hours followed by intraperitoneal injection of 2-BP (50 mg/kg body weight) or vehicle for additional 3 hours. Volcano plots of RNA-seq showing fold-change and P-value for the comparison of spleens from mice treated with 2-BP versus Veh (n = 3) with cutoffs set at FC > 2 or < 0.5, and P-value < 0.05 were shown. Representative genes associated with pyroptosis are highlighted. b, c, Eight-week-old male mice of the indicated genotypes were injected intraperitoneally with LPS (54 mg/kg body weight) for 6 hours. Volcano plots of RNA-seq showing fold-change and P-value for the comparison of spleens from Dhhc7^-/- versus Dhhc7^+/+ mice (n = 3) (b) or Apt2^-/- versus Apt2^+/+ mice (n = 3) (c) with cutoffs set at FC > 2 or < 0.5, and P-value < 0.05 were shown. Representative genes associated with pyroptosis are highlighted. d, Eight-week-old Gsdmd^-/- male mice were injected intraperitoneally with 2-BP (50 mg/kg body weight) or vehicle for 2 hours, followed by intraperitoneal infection with 10³ CFU of S. Typhimurium. The survival ratio of the mice was monitored at the desired stages. e, f, Eight-week-old male mice of the indicated genotypes were infected intraperitoneally with 10³ CFU of S. Typhimurium. The survival ratio of the mice was monitored at the desired stages. g, h, A relay model of palmitoylation and depalmitoylation that regulates GSDMD activation in pyroptosis. In the absence of DHHC7, GSDMD palmitoylation and its processing are decreased, leading to reduction of pyroptosis (g). In the absence of APT2, membrane-associated GSDMD-NT fails to undergo depalmitoylation, which reduces the oligomerization and pore-forming activity of GSDMD-NT (h). n = 6 mice for each group (d–f). Log-rank (Mantel–Cox) test (d–f). Unpaired two-tailed Student’s t-test (a–c).

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