Analysis of NETs formation. In all the panels, red bars refer to WT mice, and blue bars to APP-KO mice. Data are expressed as mean ± SEM of at least 5 different determinations. (A) NETs formation in purified neutrophils from WT and APP-KO mice was visualized by staining for DNA with Hoechst (blue) and for citrullinate Histone H3 (red). Representative images of resting and PMA-stimulated neutrophils are reported in (i), and quantification of the NETs formation expressed as percentage of neutrophils extruding NETs is reported in (ii). ***P < .005. (B) Analysis of NETs formation induced by incubation of WT and APP-KO neutrophils with WT platelets, APP-deficient platelets, or no cells (none), as indicate in the bottom. (C) Flow cytometry analysis of circulating platelet–leukocyte aggregates in blood from WT and APP-KO mice untreated (none) or on stimulation with 0.5 U/mL thrombin, as indicated. *P < .05; **P < .01. (D) Venous thrombi were isolated from WT and APP-KO mice 24 hours on IVC ligation, and stained for the NETs-related markers CRAMP and citrullinated histone H3 (Cit-H3), as indicated. (i) Some representative images, whereas (ii) reports the quantification of the positive cells counted form 5 positive stained fields. Results are reported as mean ± SEM. *P < .05; **P < .01.