CUT&Tag of human Du145 cell line: DMRT1 for 1,6 hexanediol condition 1
The CUT&Tag assay (Vazyme, TD903) was conducted according to the manufacturers instructions. Particularly, for 1,6 hexanediol condition, the cells were cultured in complete media containing 1.5% (w/v) 1,6-hexanediol (Sigma-Aldrich, 240117) for 30 min according to previous reports60. Approximately 1  105 cells were bound to ConA beads. The primary antibody (Invitrogen, PA5-29884) was incubated with the beads-cell mixture overnight at 4 C. Next, the secondary antibody (Vazyme, Ab207-01-AA) was added and incubated for 60 min at room temperature, and the pA/G-Tnp Pro was added into each sample. After extracting adaptor-ligated DNA fragments, indexing primers (TD202, Vazyme) were used to amplify the library. The amplified product was purified by VAHTS DNA Clean Beads (Vazyme, N411-01) and sequenced on the Illumina sequencing platform by Genedenovo Biotechnology Co., Ltd (Guangzhou, China).