Figure 4

Intrinsic apoptotic signals induced by MLN4924 were strengthened by P27 silencing in gastric cancer cells. AGS and MGC80-3 cells transfected with NC or siP27 were treated with DMSO or 0.3 μM MLN4924 for 72 h. (a) Sub-G1 accumulation following combination treatment. Cells were subjected to propidium iodide staining and FCM for analysis of cell cycle profile. (b) Expression of apoptotic proteins following combination treatment. Cells were subjected to immunoblotting analysis of the expression of the indicated proteins. (c) Enhanced caspase-3 cleavage by P27 silencing in MLN4924-treated cells. Cells were subjected to FITC-DEVD-FMK staining and FCM for cleaved caspase-3 quantification. For panel c, n=3. NC=Non-sense control siRNA; D=DMSO; M=MLN4924; si=siRNA for P27