The effect c-Myc siRNA transfection on c-Myc downstream molecules and cell cycle distribution. (A) Comparison of the effect of c-Myc siRNA and scrambled (nontargeting) siRNA on the expression of c-Myc downstream molecules. Transfection of CNE-2 cells with c-Myc or scrambled (nontargeting) siRNA for 48 h. Cells were then subjected to Western blotting using anti-c-Myc, -cyclin D1, -p21, and -p53 antibodies as described in Materials and Methods. Comparison of the effect of scrambled (nontargeting) siRNA (B) and c-Myc siRNA (C) on cell cycle distribution of CNE-2 cells using PI staining and flow cytometry. Each histogram is representative of three experiments. (D) Analysis of cell cycle distributions showed that, compared to srambled siRNA, c-Myc siRNA induced a conspicuous increasing of cells in S phase in CNE-2 cells at 48 h. Bar heights, average of three independent experiments; bars, SD.