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

Figure 6. Twist1 binds and promotes gene expression from an ECR within Intron1 of Semaphorin3C (Sema3C).

Figure 6

A. An ECR is located +949 to +1125 bps from the mouse Sema3C +1 site. Cross-species genomic alignment of the E-box consensus sequence (shaded grey) and mutated sequence (mutated bps indicated by *) are indicated. B. Sema3C-Intron1 or Sema3C-Intron1Mut was co-transfected into HEK293 cells with empty vector or with Twist1, E12, or Twist1 and E12 expression vectors and luciferase reporter assays performed. Fold change over the empty vector control set to 1 is shown with SEM. C. ChIP assays were performed with anti-Twist1 in mouse E12.5 ECCs and E17.5 AV valves and quantified by qPCR. Fold enrichment was evaluated by comparing anti-Twist1 IP of E12.5 ECCs or anti-Twist1 IP of E17.5 valves versus IgG (negative control) set to 1. Statistical significance was determined by Student's t-Test, p = ≤0.05 indicated by *. All experiments were performed in biological and technical triplicate. Histograms are a compilation of n = 3 experiments.