Fig. 2.
Localization of ROS production. Panels A and B. SKOV3 cells transiently expressing pHyPer-cyto, a reporter for cytoplasmic H2O2, were labeled with MitoSOX to measure mitochondrial superoxide production, before treatment with 100 mM Taxol. Taxol treatment leads to increased ROS production in mitochondria but not in the cytosol of the cell 30 min following treatment. Panel B. Changes in MitoSOX fluorescence were measured using a Zeiss LSM510 laser scanning confocal microscope with a 40× objective, and individual cells were analyzed for ROS production using LSM Image Browser Software from Zeiss. The data represent the average change in MitoSOX fluorescence between time zero and 30 min post treatment of 10–30 cells total from at least three separate time courses. Error bars represent the standard error of the mean. Statistical significance relative to control was calculated using Student's two tailed t-test assuming equal variances. Values are indicated by * p-value < 2 × 10−3, ** p-value < 7 × 10−5, ‡ p-value < 2 × 10–15. Panel C. SKOV3 cells were treated for 24 h with indicated concentrations of cisplatin, then labeled with DCF-DA for 10 min before imaging with an inverted epifluorescence microscope.