Figure 7.

UNC119Ab and KRASB WT synergistically induce apoptosis, which depends on RASSF6. A and B, HCT116 cells were transfected with pBudCGFP-SUMO (Cont), pCIneoGFP-UNC119Ab (UNC119Ab), and pCIneoMyc2-KRASB WT (KRASB WT). B, cells were transfected with control siRNA (siCont) or RASSF6 siRNA (siRASSF6#1 and #2). 24 h later, cells were replated in a 12-well plate and transfection was performed. 24 h after transfection, cells were immunostained with anti-cytochrome c (Cyt C) and anti-apoptosis-inducing factor (AIF) antibodies. Nuclei were visualized with Hoechst 33342. 50 GFP-positive cells were observed. Ratios of cells with nuclear condensation, cytochrome c release, and AIF release were calculated. The validation of RASSF6 silencing was performed by quantitative RT-PCR. Data are means with S.E. *, p < 0.05; **, p < 0.01; and ***, p < 0.001. Bar, 10 μm. Three experiments were performed. Fifty cells were evaluated for each condition. Statistical analyses were performed with one-way ANOVA with Tukey's test.