Figure 5.

CCL5 activates β-catenin pathway. (a) The expression of β-catenin was determined by real-time PCR in SW1116 cells treated with CM/TCM or 20 ng/ml CCL5 for 24 hours. *P<0.05, ***P<0.001 versus control group; (b,c) The nuclear expression of β-catenin was determined by western blot in SW1116 cells treated with CM/TCM (b) or 20 ng/ml CCL5 (c) for 24 hours. **P<0.01, ***P<0.001 versus control group. Representative blots were shown in above; (d) The effect of Wnt/β-catenin inhibitor DKK1 on TCM- or CCL5-induced migration was determined by transwell assay. 2 μM DKK1 was added into SW1116 cells and seeded in the upper chamber, TCM or 20 ng/ml CCL5 were added in the lower chamber. Quantification was presented as mean±S.D. from three independent experiments. *P<0.05, ***P<0.001; (e,f) 2 μM DKK1 was added simultaneously with 20 ng/ml CCL5 or TCM into SW1116, the expression of Slug was determined by real-time PCR (e) or western blot (f). Data are presented as the means±S.D., n=3. *P<0.05, **P<0.01, ***P<0.001. Representative blot was shown in below f