title: CUT&RUN of mus musculus: Control
design: Cut&Tag-seq was performed using the Hyperactive Universal CUT&Tag Assay kit for Illumina (Vazyme, TD904). Briefly, mouse T cells were cultured for 6 days under control or mannose condition. Then, 510^5 mouse T cells were processed by centrifugation between buffer and regent exchanges in an Ultra-low attachment tube. Cell nuclei was isolated by permeabilizing cells in NP40-Digitonin Wash Buffer and incubated with Con A beads at room temperature for 10min. Cells were resuspended in antibody buffer pre-mixed with primary antibody (anti-H3K27me3, anti-H3K4me3, or anti-IgG) and incubated on a rotator at 4C overnight. The cell nuclei was then washed and incubated with a secondary antibody at room temperature for 30-60min. To harvest DNA fragmentation, the protein A/G-Tn5 pro adapter complex was added into the cells and incubated on a rotator at room temperature for 1h. The amplified and purified DNA was used for library preparation and sequence using PE150 by Illumine Nova6000 sequencer.