Fig. 4: Genetic ablation of neutrophil GSDME enhanced efferocytosis in peritonitis.

a Experimental scheme for assessing efferocytosis of ex vivo-aged neutrophils in peritonitis. Neutrophils from WT and Gsdme KO mice were cultured for 20 h and stained with 1 μM CFSE for 10 min. CFSE-labeled aged neutrophils were intraperitoneally injected into recipient mice challenged with heat-inactivated E. coli. (HIEC, 10⁷ cfu) for 48 h. 1 h post adoptive transfer, the peritoneal cavity was lavaged to collect cells to assess engulfment of CFSE⁺ neutrophils by macrophages. b The percentage of adoptively transferred neutrophils engulfed by macrophages was assessed by flow cytometry. After being engulfed, adoptively transferred neutrophils became ly6g negative but remained CFSE positive. All data are represented as mean ± SD, n = 4 mice, **P < 0.01 (P = 0.0001). c The percentage of macrophages engulfing at least one apoptotic neutrophil was assessed by flow cytometry. In the peritoneal cavity, F4/80⁺CD11b⁺Ly6c⁻ macrophages could engulf apoptotic neutrophils. After engulfing adoptively transferred neutrophils, macrophages became CFSE positive. All data are represented as mean ± SD, n = 3 mice, *P < 0.05 (P = 0.0126). d Experimental scheme for assessing efferocytosis of in vivo-aged neutrophils in peritonitis. Fresh neutrophils from WT and Gsdme KO mice were stained with 1 μM CFSE for 10 min. CFSE-labeled fresh neutrophils were intraperitoneally injected into recipient mice challenged with HIEC for 48 h. 6 h post adoptive transfer, the peritoneal cavity was lavaged to collect cells to assess engulfment of CFSE⁺ neutrophils by macrophages. e The percentage of adoptively transferred neutrophils engulfed by macrophages was assessed by flow cytometry. All data are represented as mean ± SD, n = 7–8 mice, *P < 0.05 (P = 0.0293). f The percentage of macrophages engulfing at least one apoptotic neutrophil was assessed by flow cytometry. All data are represented as mean ± SD, n = 4 mice; **P < 0.01 (P = 0.0002). Source data are provided as a Source Data file.