Fig. 3

Heparin and DNase did not reduce NET-induced platelet aggregation. a Representative traces showing dose-dependent aggregation of washed human platelets (WP, 3 × 10⁸ /mL) in response to calf thymus histones (CTH; 1, 5, 20 and 40 μg/mL). b Representative aggregation traces showing the effect of heparin on NET and CTH-induced platelet aggregation. The reactions were allowed to proceed for 18 min, then collagen (5 μg/mL) was added to the CTH-Heparin reaction to test platelet functional response. c Representative aggregation traces showing the effect of DNase on NET-induced platelet aggregation. NETs were pre-treated with DNase (20, 200 U/mL) for 30 min at 37 °C before addition to WP (3 × 10⁸/mL). Traces are representative of 3 independent experiments. d Bar graphs depict % of platelet aggregation treated with NET alone or with Heparin and DNase. Percentage platelet aggregation was measured at 20 min. In all assays, NETs constituted 10% of final reaction volume and contains 292 ± 172 pg/mL of NET-elastase. Data are expressed as mean ± SEM,*P < .05; ns: non-significant, n ≥ 3