Figure 4.
Ki8751 treatment enhances reactive oxygen species (ROS) production of breast cancer cells. The breast cancer cells, MCF-7 and MDA-MB-231, were seeded in 6-well plates at a density of 1.5 × 105 cells/well with vehicle (0.01% dimethyl sulfoxide), 2.5, or 5 μM Ki8751. After 24, 48, and 72 h of culture, the cells were harvested and stained with 2′,7′-dichlorofluorescin diacetate (20 μg/mL) at 37 °C in the dark for 20 min. ROS fluorescence of the cancer cells were analyzed using a Beckman Coulter FC500 flow cytometer. Representative overlay histograms of MCF-7 (panel A) and MDA-MB-231 cells (B) were shown. Fold increases of ROS fluorescence are plotted as the mean ± SEM (MCF-7 cells, C; and MDA-MB-231 cells, D). *P < 0.05; **P < 0.01 as compared to the vehicle-treated controls; n = 6.