Fig 1: Effects of pre-treatment with mitochondrial inhibitors (antimycin, oligomycin) or an uncoupler (CCCP) on human platelet function in vitro. Washed human platelets were pre-treated with vehicle (control), antimycin (2 µg/mL), oligomycin (10 µg/mL) or CCCP (100 µM) for 15 min at room temperature, followed by addition of human thrombin (0.5 U/mL) for 30 min, or as indicated. (A) Representative tracings showing thrombin (0.2 U/mL)-induced aggregation (1-4) or ATP release (1’-4’) of platelets pre-treated with vehicle (1,1’), antimycin (2,2’), oligomycin (3,3’) or CCCP (4,4’). (B, C) Bar diagram quantifying mean platelet aggregation and platelet dense granule secretion in different samples, respectively. (D-F) Representative histogram overlay plots showing binding of FITC-PAC1 (BD Biosciences #340507), PE-annexin V (Biolegend #640908), and PE-anti-CD62P (BD Biosciences #550561), respectively. (G-I) Corresponding bar diagrams quantifying aIIbß3 integrin activation, phosphatidylserine exposure and P-selectin expression, respectively. Each dot represents an independent observation. Data are presented as mean ± standard error of mean. *P<0.05 with respect to vehicle-treated thrombin-stimulated platelets. #P<0.05 with respect to vehicletreated unstimulated platelets. Significance in difference of means was tested by repeated measures analysis of variance and the Dunnett multiple comparison test. PS: phosphatidylserine; Thr: thrombin.