Figure 5. Effect of Mito-TEMPO on caspase-3 activation, depolarization of ΔΨm, and PS exposure in hyperthermia-treated platelets.

(A and B) Washed platelets were incubated at indicated temperatures for 3 h (A) or pre-incubated with Mito-TEMPO and solvent control at 37°C for 15 min and then incubated at 42°C for 3 h (B). Treated platelets were lysed and analyzed by Western blot with anti-cleaved p17 fragment of caspase-3. Actin levels were assayed to demonstrate equal protein loading. Representative results of three independent experiments are presented. (C–F) Washed platelets were pre-incubated without (C and D) or with Mito-TEMPO and solvent control at 37°C for 15 min (E and F), and then incubated at the indicated temperature for 3 h. Treated platelets were incubated with TMRE (C and E), or annexin V-FITC (D and F), and analyzed by flow cytometry. Representative flow cytometric histograms are shown (C and D). 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 RT for 3 h (E and F). The percentage of RT platelets pre-incubated with solvent is presented as mean ± SEM from three independent experiments. *P<0.05 as compared with solvent control at an identical temperature.