Figure 3.

WHY1 protein binds to the WRKY53 promoter. A, Schematic depiction of the ERE (GTCAAAAT) and AAATCCC sequence motifs found in the promoter region of WRKY53. The region from −321 to −287 nucleotides upstream of the ATG codon of WRKY53 is used for EMSA. B, EMSA performed with purified protein (pp) from E. coli extracts containing recombinant 6xHis-WHY1-HA or crude extract protein from the transgenic plants oeWHY1-HA5 (oe5), the why1-1 mutant (why1), and wild-type plants (wt). Unlabeled oligonucleotides were used in excess amounts (50-fold) as competitors. C, Western blot showing the relative amounts of WHY1 protein for a loading control. An antibody against the HA tag was used to detect HA-tagged WHY1 proteins.