Figure 4.

Analysis of BdTHX1 binding site. A and B, BdTHX1 binding site was analyzed by using mutant competitors in EMSA. The sequences of mutant competitors used in EMSA (B) are shown in A. A, The binding motif is highlighted in bold, and the mutated nucleotide of each competitor is lowercased. B, EMSA was performed with biotin-labeled probe F5 and wheat germ cell-free synthesized BdTHX1. Free DNA and protein-DNA complex were separated by 5% TBE polyacrylamide gels. Excess amounts of unlabeled probes F5, M1, M2, M3, M4, M5, M6, M7, M8, and M9 (200-fold molar excess over labeled DNA) were included as competitors. C, The motif created using program MEME motif analysis of the top 100 peaks by IDR score of ChIP-seq data. The height the label of each residues represents its frequency at that position. The GT-motif nucleotides are underlined.