Fig. 4. Tead proteins bind the CE in a sequence-specific manner. (A) Gel mobility shift assay showing that Tead2 and Tead4 bind to the CE. Wild-type CE was used as a probe. Positions of the Tead-DNA complexes are indicated by arrows. Combination of protein and competitor are indicated above each lane. Sequences of the probe and competitors are described in Fig. 3A. (B) Sequences of the competitors used in panel C. Sequences of NF-kB-BS and GT-IIC were adapted from those described previously (Davidson et al., 1988; Fujita et al., 1992). Altered residues in CE/NF-kB and CE/GT-IIC are indicated in red lower case. Core recognition sequences of Rel and Tead are indicated by boxes. Position of the M4 mutation that is essential for NE activity is shown in red in the CE sequence. (C) Comparison of DNA-binding activities of Rel and Tead. Combinations of proteins and competitors are indicated above each lane. Positions of protein-DNA complexes are indicated on the right. (D) Transgenic embryos carrying a mutant enhancer in which CE was altered to CE/GT-IIC retained gene expression in the node and notochord (n=6/7). (E) Transgenic embryos carrying a mutant enhancer in which CE was altered to CE/NF-kB lost gene expression in the node/notochord (n=0/4). n, node; nc, notochord.