Fig. 2. GSDMD-dependent pyroptosis ensues in septic platelets.

a, Representative low- and high-power TEM fields showing platelet ultrastructures from patients with severe sepsis (with or without septic shock) (n = 5) and HS (n = 5). Red arrowheads indicate swelling and rupture of the platelet plasma membrane during severe sepsis. The magnified TEM showed a swollen platelet that had ruptured (region around red arrow) and released its cellular contents. Scale bars, 1 μm and 500 nm. b, Colocalization of CD41 (green), ASC (red) and NLRP3 (blue) in platelets from severe sepsis (with or without septic shock) (HS, n = 8; severe sepsis, n = 8). Purple indicates overlap. Scale bars, 5 μm and 1 μm. c, Expression and quantification of pro-caspase 1 and activated caspase 1 in platelets from severe sepsis (with or without septic shock) (HS, n = 6; severe sepsis, n = 6). d, Expression and quantification of GSDMD and GSDMD N terminus proteins in platelets from severe sepsis (with or without septic shock) (HS, n = 6; severe sepsis, n = 6). e, FACS analysis of the activated caspase 1 in platelets from severe sepsis (with or without septic shock) (HS, n = 13; severe sepsis, n = 23). f, Expression of IL-1β in platelets from HS and sepsis groups (HS, n = 8; severe sepsis, n = 8). g, Live-imaging of platelets from Gsdmd^fl/fl PF4-Cre mice and Gsdmd^fl/fl mice induced by LPS (10 μg ml⁻¹) and NIG (5 μM) for 20 min (n = 6). Scale bars, 10 μm. h, Representative TEM field of platelets from Gsdmd^fl/fl PF4-Cre mice and Gsdmd^fl/fl mice with or without CLP operation (n = 6). Scale bar, 1 μm. Data were presented as mean ± s.d. or median with interquartile range. Statistical analysis was conducted using unpaired two-tailed t-test (b–g), and Kruskal–Wallis test and Dunn’s multiple comparisons test (h). Sham, sham-operated mice.

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