Figure 2. Hyperthermia increase mitochondrial superoxide production.

(A and B) MitoSOX^TM Red-loaded platelets were incubated at the indicated temperatures for 3 h (A) or1-3 h (B). As a positive control, loaded platelets were incubated with antimycin A at 37°C for 30 min. Samples were then analyzed by flow cytometry. Representative flow cytometric histogram is shown (A). Data are expressed as a percentage of platelets which were incubated at RT for 1h (B). Percentage of RT 1h is presented as mean ± SEM from three independent experiments. *P<0.017 (after a Bonferroni correction) compared with RT 1 h, **P<0.017 (after Bonferroni correction) as compared with RT 2 h, #P<0.017 (after Bonferroni correction) as compared with RT 3 h. (C) MitoSOX^TM Red-loaded platelets were pre-incubated with apocynin, Mito-TEMPO, L-NAME, ETYA, or solvent control at 37°C for 15 min, and then incubated for 3 h at 40°C or 42°C, and further analyzed by flow cytometry. Data are expressed as a percentage of platelets that were pre-incubated with solvent control at 37°C for 15 min and then incubated at 40°C for 3 h. Percentage of 40°C platelets pre-incubated with solvent control is presented as mean ± SEM from three independent experiments. *P<0.013 (after Bonferroni correction) as compared with solvent control at 40°C, **P<0.013 (after Bonferroni correction) as compared with solvent control at 42°C. Antimycin A is labeled as Ant.