To characterize the potential binding targets of OsDOF3, we conducted the CUT&Tag assay in rice protoplasts. Transfected protoplasts were subjected to fluorescence-activated nuclei sorting, and a minimum of 100,000 GFP-positive nuclei were collected per biological replicate. we conducted a CUT&Tag assay inplants. Two-week-old seedlings were harvested, and leaf sheath tissues were collected for nuclear isolation. Tissues were finely minced in 20 mL of ice-cold Lysis Buffer (15 mM Tris-HCl [pH 7.5], 20 mM NaCl, 80 mM KCl, 0.5 mM spermine, 0.2% Triton X-100, 5 mM β-mercaptoethanol, and 1× protease inhibitor cocktail) using a surgical blade. The homogenate was filtered through a 40-μm cell strainer, and nuclei were stained with 1 μg/mL DAPI (4',6-diamidino-2-phenylindole) for fluorescence-activated nuclei sorting. Flow cytometric analysis and sorting were performed using a Sony SH800 Cell Sorter equipped with a 405-nm laser. A minimum of 100,000 nuclei were collected per biological replicate. The sorted nuclei were immediately processed for chromatin enrichment and library preparation steps using the CUT&Tag Assay Kit (Vazyme, TD904-C2) according to the manufacturer's instructions.