Figure 6

Twist1 binding region sequences are associated with genes in the Wnt signaling pathway. A. Wnt pathway genes associated with Twist1 E-box-containing binding regions detected by ChIP-seq in E12.5 ECCs include receptor tyrosine kinase-like orphan receptor 2 (Ror2-b), homeodomain interacting protein kinase 2 (Hipk2), secreted frizzled-related protein 1 (Sfrp1), frizzled homolog 4 (fzd4), and frizzled homolog 8 (fzd8). Twist1 binding to these sequences was confirmed by ChIP with anti-Twist1 in E12.5 ECCs followed by quantification by qPCR. B. Wnt pathway genes associated with Twist1 E-box containing binding regions detected by ChIP-seq in E10.5 limb buds include Wnt5b, lymphoid enhancer binding factor 1 (Lef1), nemo like kinase (Nlk), casein kinase 1, epsilon (Csnk1e), disabled 2 interacting protein (Dab2ip), mutated in colorectal cancers (Mcc), and frizzled homolog 3 (Fzd3). Twist1 binding to these sequences was confirmed by ChIP with anti-Twist1 in E10.5 limb bud followed by quantification by qPCR. All ChIP experiments were quantified with qPCR, and fold-enrichment was calculated relative to normal rabbit IgG control. All analyses were performed in biological triplicate. * indicates p < 0.05 as determined by student’s t-test comparing IgG and Twist1 ChIP qPCR ΔΔCt values.