Figure 6. Inhibition of PAD4 dampens thromboinflammation by decreased neutrophil infiltration, neutrophil H3Cit expression and reduced endothelial activation.

(A) Representative immunostainings of LV sections and quantification of double-positive (Ly6G⁺ [red] and H3cit⁺ [green]) cells indicative of activated neutrophils in the myocardium of mice with collagen-induced arthritis (CIA) treated with the vehicle and mice with CIA treated with the PAD4 inhibitor, respectively (scale bar: 50 μm) (n = 7–8). (B) Quantification of infiltrating CD45⁺ and LY6G⁺ cells in the myocardial tissue of mice with CIA treated with the vehicle and mice with CIA treated with the PAD4 inhibitor, respectively, assessed by flow cytometry (n = 8). (C) Representative LV sections stained with DAPI⁺ (blue) and VWF⁺ (green). Quantification of VWF deposition in the myocardium of mice with CIA treated with the vehicle and mice with CIA treated with the PAD4 inhibitor (scale bar: 200 μm). (D) Myocardial tissue levels of IL-1β in mice with CIA treated with the vehicle and mice with CIA treated with the PAD4 inhibitor. Data are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001; unpaired T test. (B) Wilcoxon matched pairs signed rank test.