Extended Data Fig. 10. Immunofluorescence of pyroptotic platelets in NETs and the change of platelet counts in Gsdmdfl/fl PF4-Cre mice by CLP.
a, PMNs (S100a9-/- or WT) were incubated with 50 nM PMA to induced NET formation for 4 hours, and then incubated with platelets for another 4 hours. Representative immunofluorescence of PMNs incubated with platelets. Cells were stained with Hoechst for DNA (blue), anti-MPO for PMNs or NETs (cyan), CD41 for platelet (red) and activated caspase 1 for pyroptosis (green); scale bars: 25 μm and 5 μm. b, In the CLP-induced sepsis model, platelet counts in Gsdmdfl/fl PF4-Cre mice and littermate control Gsdmdfl/fl mice were assessed at 0, 2, 4, and 6 hours using a hematology analyzer (n = 5). Data was presented as mean ± SD. Two-way ANOVA and Tukey’s multiple comparisons test for b. Abbreviation is as follow: PLT, platelet; NS, not statistically significant; PMNs, polymorphonuclear neutrophils; PMA, phorbol myristate acetate; MPO, myeloperoxidase; Sham, sham-operated mice; CLP, CLP-induced sepsis mice; GSDMD, Gasdermin D.