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. 2011 Dec 29;6(12):e29758. doi: 10.1371/journal.pone.0029758

Figure 7. Twist1 binds and promotes gene expression from ECRs associated with Gadd45a and Rab39b.

Figure 7

A. An E-box containing ECR is located −3240 to −3456 bps from the mouse Gadd45a +1 site with the E-box sequence indicated by grey shading. B. (left) Gadd45a-promoter plasmid was co-transfected into HEK293 cells with empty vector or with Twist1, E12, or Twist1 and E12 expression vectors and luciferase reporter assays performed (left). Fold change over the empty vector control set to 1 is shown with SEM. ChIP assays were performed with anti-Twist1 in mouse E12.5 ECCs and mE17.5 AV valves versus IgG (negative control) set to 1 (right). C. An E-box containing ECR is located within the mRab39b3′UTR, +5891 to +6093 bps from mRab39b +1 site with the E-box indicated by grey shading. D. Rab39b-3′UTR was co-transfected into HEK293 cells with empty vector or with Twist1, E12, or Twist1 and E12 expression vectors and luciferase reporter assays performed (left). Fold change over the empty vector control set to 1 is shown with SEM. ChIP assays were performed with anti-Twist1 in mouse E12.5 ECCs and E17.5 AV valves versus IgG (negative control) set to 1 (right). Statistical significance was determined by Student's t-Test, p = ≤0.05 indicated by *. All experiments were performed in biological triplicate. Histograms are a compilation of n = 3 experiments.