Figure 3

HOXB13 suppresses expression of TCF4 at the post-transcriptional level. (A) Cells were infected with either Ad-GFP or Ad-GFP-HOXB13 virus (100 MOI) for 48 h. Nuclear proteins were separated by gel electrophoresis followed by Western blot analysis. (B) Potential promoter region of TCF4 was cloned into pGL-luc system (pGL-TCFp-luc) followed by transient transfection assay. (C) Cells were transfected with pGL-TCF4p-luc and either pFLAG or pFLAG-HOXB13. ^*P>0.2; ^**P<0.05. (D) HCT116 cells were infected with Ad-GFP or Ad-GFP-HOXB13 (100 MOI) followed by extraction of total RNA. Real-time RT–PCR was performed for TCF4 expression and evaluated by standard curve methods. RNA from normal rectum was used to draw the standard curve for TCF4 and β-actin. ^*P=0.32.