Figure 6

Tumorigenicity of ATF2 in vivo. B16F10 cells stably transfected with ATF2^T52A or ATF2 shRNA were injected into C57/BL6 mice and the tumor volume for ATF2^T52A tumor xenografts (A) and ATF2 shRNA tumor xenografts (B) was measured on the indicated day. Bars indicate s.e.m. *P < 0.01, Student’s t test. (C) The ATF2 mRNA levels in the cancer tissues isolated from tumor xenograft mice were measured by quantitative real-time RT-PCR (ANOVA, *P < 0.01, n = 6). (D) Representative pictures of mice bearing tumors at 3 weeks after B16F10 cells injection. (E) Mitochondrial and nuclear fractions were prepared from tumor xenografts and subjected to western blotting with an anti-ATF2 antibody at 3 weeks after B16F10 cells injection. The morphometry of ATF2 are indicated at right. The columns provide average values of at least 3 independent experiments performed in triplicate; the bars, SE. *P < 0.01 compared with the corresponding control. (F) After injection of the indicated tumor cells for 6 days, the xenografts were collected, and the in situ tumor cells were harvested and subjected to FACS apoptosis analyses by Annexin V/PI staining. Diagram presented quantitative analysis of three similar experiments. *P < 0.01, ANOVA. Data are means ± s.e.m.