FIGURE 7.

Synergistic anticancer effect of Emi1 knockdown in combination with doxorubucin in normal cells. A, Emi1 or control siRNA was transfected into NHDF cells, and cells were collected after 48 h. The indicated proteins in siRNA-treated NHDF cells were examined by Western blotting. B, Emi1 siRNA and/or cyclin A siRNA were transfected into NHDF cells. The left panel shows the expression of Emi1 and cyclin A examined by Western blot analysis after 48 h of siRNA transfection. β-Actin expression was used as a loading control. The right panel shows DAPI staining and percentage of cells with polyploidy after Emi1 siRNA and/or cyclin A siRNA transfection. Cells were stained with DAPI to visualize the nuclei, and percentage of cells with polyploidy was determined by DNA content analysis after PI staining using a flow cytometer. *, p < 0.05. C, flow cytometric analysis of annexin V and PI staining in control and Emi1 siRNA treated NHDF or HFL III cells after treatment with doxorubucin (DOXY; 0.5 μg/ml) for 12 h. We performed three independent experiments.