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

Figure 2. Twist1 binds and promotes gene expression from an ECR within Intron1 of Cadherin-11 (Cdh11-Intron1).

Figure 2

A. An E-box-containing ECR is located +36256 to +36552 bps from the chicken Cdh11 transcriptional start site (+1). Cross-species genomic alignment of the E-box consensus sequence (shaded grey) and mutated sequence (mutated bps indicated by *) are indicated. B. Cdh11-Intron1 or Cdh11-Intron1Mut plasmids were 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 mE12.5 ECCs and mE17.5 AV valves and quantified by qPCR. Fold enrichment was evaluated by comparing anti-Twist1 IP of mE12.5 ECCs or anti-Twist1 IP of mE17.5 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.