Fig. 3
H3K4me3 and histone H3 distribution patterns at RD29B and RD29A genes in response to dehydration stress and to treatment with JA. The levels of histone H3K4me3 modifications and of histone H3 determined by ChIP–qPCR assays with specific antibodies under various treatments are annotated as indicated in Fig. 1B. A H3K4me3 levels at three different locations along the RD29B gene indicated by the gene diagram on top. H3K4me3 data were normalized versus values for histone H3 at the same locations. H3K4me3 levels before and after JA-treatments are shown in the same scale. B Histone H3 distribution determined by ChIP–qPCR with histone H3-specific antibodies and DNA recovered from the regions indicated in the gene diagram on top. ChIP–qPCR data from JA-treatments are shown in the same scale as those from untreated samples. ChIP–qPCR values obtained for IgGs at the specific amplicons are shown below; C H3K4me3 levels at RD29A at the regions indicated by the gene diagram on top. H3K4me3 data were normalized versus values for histone H3 at the same locations. Data from JA-treated and JA-untreated samples are shown in the same scale; D Histone H3 distribution determined by ChIP–qPCR with histone H3-specific antibodies and DNA recovered from the RD29A regions indicated in the gene diagram on top. ChIP–qPCR values obtained for IgGs at the specific amplicons are shown below. Numbers below bars indicate probed regions, as illustrated by the schematic diagram of the genes on top: promoter regions (1), untranslated regions (gray box), exons (dark box), and introns (thin lines between exons); region 2 is immediately downstream of TSS where the peak of K4me3 accumulates, and region 3 corresponds to downstream 3′-end sequences. Experiments were repeated three times, each with 3 RT-qPCR measurements, and the representative experiment shown indicates the mean ± SEM, n = 3 replicates. Different letters above bars indicate significant difference among the treatments in the region of interests (p < 0.05 according to Tukey’s multiple range test)