Figure 2
Survivin expression increased mRNA and protein levels of β-catenin/Tcf-Lef target genes: A,B: HEK293T cells (5 × 105) were seeded in 6-well plates and transfected with pEGFP-C1 or pEGFP-survivin (1.5 μg). A: After 24 h Cyclin D1, Runx-2, COX-2 and survivin mRNA levels were assessed by semi-quantitative RT-PCR. Actin was used as an internal control B: Runx-2,COX-2 and Cyclin D1 mRNA levels were assessed by qPCR. Ribosomal 18S RNA was used as an internal control. Relative Units (RU) reflect normalization to controls. Values are means of results obtained in three independent experiments. Statistically significant differences compared to mock-transfected controls are indicated (* p < 0.05). C: Cyclin D1, COX-2 and endogenous survivin levels were evaluated by western blotting. Actin was used as an internal control. Expression of GFP and GFP-survivin in the respective lanes was revealed with a GFP-specific antibody. D, E: Wild type MKN45 gastric cancer cells (5×105) were seeded in 6-well plates and transfected with with pEGFP-C1 or pEGFP-survivin (1.5 μg). After 24 h mRNA and protein levels were assessed by qPCR and western blotting, respectively. D: Runx-2, COX-2 and cyclin D1 mRNA levels were assessed by qPCR. Ribosomal 18S RNA was used as an internal control. Relative Units (RU) reflect normalization to controls. Values are means of results obtained in three independent experiments. Statistically significant differences compared to mock-transfected controls are indicated (* p < 0.05). E: Cyclin D1, COX-2 and endogenous survivin protein levels were evaluated by western blotting, quantified by scanning densitometry of immunoblots and normalized to actin. Numerical data shown are means of results obtained in three independent experiments. Statistically significant differences compared to mock controls are indicated (* p < 0.05).