Fig. 6

Twist1 negatively regulated Trabid expression in HCC. a The nucleotide sequences and positions of four putative Twist1 binding site in the Trabid proximal promoter are shown. Luciferase activity assays of the pGL3-Trabid promoter-reporter. b1 Serially truncated and mutation analysis b2 identified twist1-bindinf sites in the Trabid promoter. ChIP assay demonstrated the direct binding of Twist1 to the Trabid promoter in HCC cells (b3) and human HCC tissues (b4). Hepatocytes were separated from liver tissues of healthy controls (HC) and HCC patients. c ChIP assays in clinical HCC tumor samples (case #1 and case #2) and HCC cell lines. d Indicated HCC cells were transduced with or without Ad-Twist1. The levels of Twist1 and Trabid were determined using Western blot