Bmi-1 knockdown increases cisplatin-mediated ROS production in ovarian cancer cells.

Ovarian cancer cells transfected with scrambled control or Bmi-1 siRNA were treated with or without cisplatin for 24 h. Subsequently the cells were incubated with 5 µM carboxy-H2DCFDA in fresh HBSS for 30 min at 37°C. The cells were harvested with trypsin and fluorescence of the labeled cells was measured at an excitation wavelength of 485 nm and emission wavelength of 530 nm by using Fluorolog 3 (Jobin-Yvon Horiba). Ratio of mean fluorescence intensity (MFI) with respect to the untreated scrambled control is represented.