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. 2024 Jan 9;15:386. doi: 10.1038/s41467-023-44669-y

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Cleavage of GSDME during neutrophil spontaneous death. Bone marrow-derived neutrophils from WT mice were used. Results are representative of at least three biological replicates. All data are represented as mean ± SD, n = 3 independent repeats. b Representative images of neutrophils undergoing apoptosis and lytic cell death. Results are representative of at least three independent experiments. c Representative images of aging neutrophils at the indicated time points. The white arrow heads indicate swollen or “puffed” cells (lytic cell death). PI propidium iodide, BF bright field. Results are representative of at least three independent experiments. d The percentage and number of swollen, AV-positive, and PI-positive cells. All data are represented as mean ± SD. n = 4 independent repeats, *P < 0.05, **P < 0.01, ns non-significant (Percentage of AV⁺ cells: 4 h, P = 0.0273; 8 h, P = 0.0285; 12 h, P = 0.0011; 16 h, P = 0.001; 20 h, P = 0.0004; 24 h, P = 0.0067. Percentage of PI⁺ cells: 8 h, P = 0.0297; 12 h, P = 0.0011; 16 h, P = 0.022; 20 h, P = 0.0168; 24 h, P = 0.0037. AV⁺ cell number: 4 h, P = 0.0184; 8 h, P = 0.0261; 12 h, P = 0.0001; 16 h, P = 0.0001; 20 h, P = 0.0001; 24 h, P = 0.0001. PI⁺ cell number: 8 h, P = 0.0046; 12 h, P = 0.0099; 24 h, P = 0.0012). e The number of intact neutrophils remaining. All data are represented as mean ± SD. n = 3 independent repeats, *P < 0.05, **P < 0.01 (Remaining cell number: 12 h, P = 0.0119; 16 h, P = 0.024; 20 h, P = 0.0001; 24 h, P = 0.0001. Disappeared cell number: 12 h, P = 0.0132; 16 h, P = 0.0207; 20 h, P = 0.0247). f The percentage and number of healthy neutrophils. All data are represented as mean ± SD. n = 4 independent repeats, *P < 0.05, **P < 0.01, ns non-significant (Percentage of healthy cells: 8 h, P = 0.0499; 12 h, P = 0.0115; 16 h, P = 0.0001; 20 h, P = 0.0005; 24 h, P = 0.0008). g The absolute number of healthy neutrophils and neutrophils undergoing apoptosis or lytic cell death. All data are represented as mean ± SD (n = 4 independent repeats). h LDH release from cell culture supernatants. Data are mean ± SD. n = 3 independent repeats, *P < 0.05, **P < 0.01 (4 h, P = 0.042; 8 h, P = 0.0001; 12 h, P = 0.0003; 16 h, P = 0.0023). Statistical significance was examined by unpaired two-sided Student’s t test (d, e, f, h). Source data are provided as a Source Data file.