Figure 8. ZNF281 negatively regulated β-TrCP2 transcription in CRC.

(A) Schematic diagram of the β-TrCP2 proximal promoter reporter. The nucleotide positions and sequences of two putative ZNF281 binding sites are shown. Luciferase activity of the indicated pGL3-β-TrCP2 promoter-reporter constructs after transfection into HEK293T cells are shown. (B) ChIP assay was performed using a specific anti-ZNF281 antibody. (C) CRC cells were transduced with a Sh-control or Sh-ZNF281 for 24 h. And the levels of ZNF281 and β-TrCP2 protein were determined using Western blot analysis. (D) Schematic of GSK3β-dependent ZNF281protein ubiquitylation and degradation byβ-TrCP2. *P <0.05, **P <0.01.