Figure 3.

In vitro effect of ENO1 overexpression or silencing on cell growth and proliferation. (A) A2780CP20 cells were stably transfected with an empty vector (EV) or with an ENO1-containing vector. Western blot analysis was performed with 50 µg of protein extracts. (B) EV and ENO1 clones (3 × 10⁴ cell/ml) were exposed to different concentrations of cisplatin for 72 h. Cell viability values were calculated relative to untreated cells. (C, D) Percentages of clonogenicity were calculated relative to EV cells. (E) A2780 cells (3 × 10⁴ cells/ml) were transiently transfected with a negative control siRNA (NC-siRNA) or the two ENO1-targeted siRNAs. Western blot was performed with 50 µg of protein extracts. (F) Densitometry analysis of band intensities from (E). Protein levels were calculated relative to NT cells. (G) A2780 cells (3 × 10⁴ cells/ml) cells were plated in 96-wells and the next day cells were transfected with different concentrations of siRNAs, as described in (E) Twenty-four hours after siRNA transfection, cisplatin (2 mM, final concentration) was added to the cells. Forty-eight hours later, cell viability was measured. Averages ± SEM are shown for three independent experiments. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.