Figure 4. RUNX3 overexpression promoted cell growth and inhibited serum starvation induced apoptosis.

A: Generation of RUNX3 overexpressing cells. The RUNX3/pcDNA3 plasmid or the vector alone was introduced into HSC3 cells, and the stable pool clones were obtained by G418 selection for 2 weeks. Exogenous expression of RUNX3 mRNA and protein was examined by RT-PCR and Western blot analysis (WB). Cul1 was used as a loading control. B: The graph shows cell growth of RUNX3 overexpressing and control HSC3 cells. Cells were plated on 24 well plates, and trypsinized cells were counted by Cell Counter (Coulter Z1) at 0, 2, 4 and 6 day. C: RUNX3 overexpression inhibited the serum starvation induced apoptosis. Cells were incubated for 0, 48 and 96 hours after serum starvation and fixed in 70% ethanol. Cell cycle distribution was determined by DNA content analysis after propidium iodide staining using a flow cytometer. For each sample, 20,000 events were stored. We performed two independent experiments. D: Flow cytometric analysis of Annexin V and propidium iodide staining in control and RUNX3 overexpressing cells after serum starvation for 96 h. We performed two independent experiments.