Figure 8.

MYC2 directly binds to the promoters of VTC1 and GSH1. A, Schematic diagram of the potential G-box and G-box-like motifs in the promoter sequences of the VTC1 and GSH1 genes. Numbers indicate the nucleotide positions relative to their corresponding translational start site (ATG), which is shown as +1. B, ChIP-qPCR analyses showing the in vivo interaction of MYC2 with the DNA fragments in the promoters of VTC1 and GSH1. The protein/DNA complexes isolated from 2-week-old MYC2-OE plants were immunoprecipitated with or without the anti-MYC2 antibodies. For each promoter, three DNA fragments (P1, P2, and P3) were used to determine the enrichment of the DNA fragment containing the G-box and G-box-like motifs. The ACTIN2 promoter fragment was used as a negative control. The data are means ± sd (n = 3 independent experiments). C, EMSA analyses showing the binding of MYC2 to the DNA probes of VTC1 and GSH1 promoters in vitro. Biotin-labeled probes were incubated with nucleoproteins extracted from 2-week-old MYC2-OE seedlings. The free and bound DNAs (arrows) were separated in an acrylamide gel. The excess cold, unlabeled probes were used as competitors (lanes 3, 4, and 5), and the mutated probes (lane 6) were produced by replacing the G-box-like motifs.