Figure 3
Survivin downregulation decreased β-catenin protein levels and β-catenin/Tcf-Lef transcriptional activity: A,B: B16F10 cells were stably transfected with 2 different shRNA targeting mouse survivin (shSUR1 and shSUR2) or luciferase (shLUC) as a control. Batch populations of cells (5×105) were seeded in 6-wells plates A: After 24 h, β-catenin, COX-2, Cyclin D1, survivin, and actin protein levels were evaluated by western blotting. Protein levels were quantified by scanning densitometric analysis of western blots and normalized to actin. Numerical data shown are the means of results obtained in three independent experiments. Statistically significant differences compared to shLUC controls are indicated (* p < 0.05). B: B16F10 (shLUC, shSUR1 and shSUR2) cells were transiently transfected with the reporter plasmids pTOP-FLASH or pFOP-FLASH (1 μg). Luciferase activity was obtained by calculating the pTOP-FLASH/pFOP-FLASH activity ratios for each condition. Data were previously normalized to values for β-galactosidase activity. Numerical data shown are the means ± s.e.m. of results obtained in three independent experiments. Statistically significant differences between shSUR1-2 and shLUC controls are indicated (* p < 0.05).