Fig. 7: SRT1, SRT2, and HDA705 have histone de-β-hydroxybutyrylase activities.

a Histone H3K9bhb levels in hda701, hda704, hda705, hda716, srt1, and srt2 CRISPR/Cas9 plants compared to wild-type (WT) plants. b In vitro de-β-hydroxybutyrylatase activity assay. Histone H3 was used as a loading control. For figures (a, b), histones were acid-extracted from the rice leaves and immunoblotted with the indicated antibodies. Images shown are representative of two independent experiments. The immunoblot signals were quantified using ImageJ. Relative quantified signals for each band are indicated, with the first WT/CK loading set as 1.00. c Ridge plots of H3K9bhb levels in hda705 and WT plants. The read counts were normalized using the DESeq2 size factor normalization method. d Cumulative density plots of H3K9bhb levels in hda705 and WT plants. P-value was calculated from a two-sample Kolmogorov–Smirnov test. The read counts were normalized using the DESeq2 size factor normalization method. e Volcano plots of differential H3K9bhb levels in hda705 relative to WT. f GO pathway analysis of the genes (n = 2971) with significantly upregulated H3K9bhb levels in hda705 in comparison to WT. g Assay of expression and H3K9bhb levels of genes in WT and hda705 rice plants. Upper: H3K9bhb ChIP-qPCR assays of chromatin isolated from CK and hda705 rice plants. Lower: RT-qPCR analysis of transcript levels in four selected genes in WT and hda705 plants. Bars indicate means ± SD from three replicates. P-value was calculated by a two-tailed, paired Student t test. Source data are provided as a Source Data file.