Research Paper Volume 12, Issue 23 pp 24424—24440

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 (I–K). 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.