Figure 5

tBHP induces the translocation of phosphatidylserine to the outer leaflet of the cellular membrane. (Top panel) After treatment with 0, 50 and 100 μM tBHP, for 1 hour, H9c2 cell were labeled with Annexin V and propidium iodide. Left panels: DIC images of H9c2 cells, treated with 0, 50 and 100 μM tBHP. Right panels: corresponding epifluorescence microscopy images of propidium iodide (red) and Annexin V (green). Images are representative from 3 different experiments. All images are the same magnification; scale bar = 20 μm. (Bottom panel) Statistical analysis of the population of control cells or cells treated with increasing concentrations of tBHP, showing the relation between treatment and annexin V/propidium iodide labeling. Analysis was performed by counting cells without labeling (healthy cells), annexin V labeling (apoptotic cells) and PI/Annexin V double labeling (necrotic cells). Data represent the mean ± SEM of 3 different cell preparations.