Fig. 3

CD133-positive HCC cells have an efficient ROS defense system. a Western blot analysis of CD133 after sorting of CD133-negative and CD133-positive HCC from Huh7 cells. b Expression of CSC-related cell surface markers (EpCAM, CD90, CD44, CD24, AFP) were examined in CD133-negative HCC and CD133-positive HCC by western blotting. c ROS accumulation in CD133-negative and CD133-positive HCC was measured after treatment with 200 μM H₂O₂ for 20 min. The data shown were from three independent experiments relative to the value for control. d CD133-negative and CD133-positive HCC were treated with the indicated concentrations of MTX, cisplatin, and As₂O₃. To examine the effect of radiation, cells were treated with 10 Gy radiation. For ROS level measurement, cells were stained with CM-H₂DCFDA and analyzed by the HCS system after 12 h treatment with anti-cancer drugs and radiation treatment