Figure 4. Inhibition of NCF2/p67phox in HCT116 cells decreases ROS and induces apoptosis. (A) HCT116 cells were transfected with either a control siRNA (si-scr) or specific NCF2/p67phox-targeting siRNA (si-NCF2/p67phox) and collected after 48 h. NCF2/p67phox protein and transcript levels were examined by western blot and real-time PCR, respectively. The western blot shown is one representative experiment. Real-time PCR shows the mean of three independent experiments. Actin is shown as a loading control. (B) ROS levels were assayed using a DCFDA staining and FACS analysis. Left panel shows one representative experiment of four independent ones. Right panel shows the mean of four independent experiments as a percentage respect to the control (presented as 100%). (C) Apoptosis levels were assayed by propidium iodide staining and FACS analysis. Percentage of sub-G₁ events (M1) is shown. Left panel shows one representative experiment of three. Right panel shows the mean of three independent experiments.