Gsdmd deficiency reduces NETs in both kidney arteries and tissue necroinflammation in focal TMA. (A) Representative immunofluorescent images of NETs (NETs), identified as CitH3-positive areas (green) originating from Ly6G-positive neutrophils (cyan), within αSMA-positive arteries (red) in the TMA kidney. DNA was visualized using DAPI (blue). (B) Representative immunofluorescent images of Ly6G-positive neutrophils (red) and CD41-positive platelets (cyan) within the αSMA-positive arteries (green) in TMA kidneys. DNA was visualized using DAPI (blue). (C-F) Quantification of CitH3 (C), Ly6G (D), CD41 (E), and Ly6G and CD41 (F)-positive areas within αSMA-positive arteries in TMA kidneys from WT and knockout (Gsdmd^−/−) mice (n = 3-4 per group). (G) Representative immunofluorescent images of NETs in the peri-infarct of TMA kidneys, identified as CitH3-positive area (green) originating from Ly6G-positive neutrophils (red). DNA was visualized with DAPI (blue). (H) Quantification of the CitH3-positive area in the peri-infarct of sham (n = 4) and TMA kidneys from WT (n = 8) and Gsdmd^−/− (n = 8) mice. (I) Absolute numbers of NETing neutrophils (CD45⁺ CD11b⁺ Ly6G⁺ CitH3⁺) in sham (n = 4) and TMA kidneys from WT (n = 8) and Gsdmd^−/− (n = 8) mice, as determined by flow cytometry. Scale bars: for panels A-B, 20 μm, for panel G, 50 μm (low magnification), and 20 μm (high magnification). The data represent mean ± SD. ∗P < .05 using unpaired Student t test for panels C-F or 1-way ANOVA with Tukey post hoc test for panels H-I.