Research Paper Advance Articles pp 24424—24440

FOXM1/DVL2/Snail axis drives metastasis and chemoresistance of colorectal cancer

Figure 6. FOXM1 bound to DVL2 and enhanced its nuclear translocation. Western blotting for the nuclear DVL2 and FOXM1 expressions in HCT-8 cells transfected with pcDNA3.1 or pcDNA3.1-FOXM1 (A), HCT-8/L-OHP and HCT-8/VCR cells transfected with shNC or shFOXM1 (B, C), HCT-8 cells treated with or without 50 ng/ml LMB for 12 h in presence of shNC or shFOXM1 transfection for 72 h (D), as indicated. Dual-luciferase reporter assay for TOPflash and FOPflash luciferase activity in HCT-8 cells transfected with pcDNA3.1, pcDNA3.1-FOXM1, or pcDNA3.1-FOXM1 plus shDVL2 for 48 h (E), HCT-8/L-OHP cells transfected with shNC, shDVL2, or shDVL2 plus pcDNA3.1-FOXM1 for 48 h (F). The relative luciferase activity was normalized against Renilla reporter pRL-SV40 activity. Coimmunoprecipitation (Co-IP) of endogenous FOXM1 and DVL2 in HCT-8/L-OHP cells and HCT-8/VCR cells (G, H). Co-IP of fusion protein HA-FOXM1 and Flag-DVL2 in total cellular lysates (TCL) and nuclear fractions in HCT-8 cells transfected with pcDNA3.1-HA-FOXM1 and/or pcDNA3.1-Flag-DVL2 for 72 h (IK). In each case, the blot is representative of immunoblots resulting from three separate experiments. Data are expressed as mean ± SD of three independent experiments. *P < 0.05.