KCTD1 Suppresses Canonical Wnt Signaling Pathway by Enhancing β-catenin Degradation
Figure 9
Effects of APC and p53 on KCTD1-mediated β-catenin degradation.
(A) HEK293 cells were transfected with TOPFLASH reporter plasmid either alone or with pCMV-Myc-KCTD1 or with pCMV-Myc-APC truncations or with both pCMV-Myc-KCTD1 and pCMV-Myc-APC truncations. (B) HeLa cells were transfected with either pCMV-Myc-β-catenin alone or with pCMV-Myc-KCTD1 or with pCMV-Myc-APC trucations or in combination as indicated. 24 h after transfection, cell lysates were detected by Western blots with mouse monoclonal anti-Myc antibodies. GAPDH was used as a loading control. (C) HEK293 cells were transfected with TOPFLASH reporter plasmid either alone or with pCMV-Myc-KCTD1 or with pCMV-HA-p53 or with both pCMV-Myc-KCTD1 and pCMV-HA-p53. (D) HeLa cells were transfected with either pCMV-Myc-β-catenin alone or with pCMV-Myc-KCTD1 or with pCMV-HA-p53 or in combination as indicated for 24 h, cell lysates were detected by immunoblotting with mouse monoclonal antibodies against Myc-tag and HA-tag. GAPDH was used as the internal control. Relative luciferase activities represent mean ±SD from at least three independent experiments after normalization to β-galactosidase activities. *, P<0.05 and **, P<0.01 compared with controls.