Fig. 5

GHD8 binds to the promoter of Ghd7. a A schematic diagram of the Ghd7 gene model showing the locations of two CCAAT-box motifs and one CORE1-like motif from the start codon ATG. The relative positions of the fragments from cp1 to cp9 used for the ChIP assay are given below the gene model; F1 indicates the fragment covering the CCAAT-box motif for the gel shift assays. b ChIP analysis of GHD8-GFP fusion protein using an anti-GFP antibody. The precipitated chromatin fragments were analyzed using the 9 primer sets for the target regions (cp1 to cp9) shown in (a). c Gel shift assays. Recombinant HD1 protein was incubated with biotin-F1 in the absence or presence of 25- or 100-fold molar excess of the unlabeled probe as the competitor. The arrow indicates the shifted band of HD1 protein. Maltose-binding protein (MBP) and His (polyhistidine)-tag protein were used as a negative control. d GHD8 interacts HD1 to binding F1 probe. e Interaction of the complex GHD8-OsHAP5b-HD1 (as shown by arrow) and F1 probe in the absence or presence of 50- or 100-fold molar excess of the unlabeled probe. The mutant F1 (MF1 or MF2) probe lacking the CCAAT box as a negative control