Cut-Tag of NCM 460 cell: transfected with plasmid SlyA
CUT&Tag experiments were conducted following the manufacturer's instructions using the Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903). SlyA protein with a 3  Flag tag (pECMV-3  Flag-N-SlyA) was heterologously expressed in NCM460 cells. 24 hours post-transfection, the culture medium was removed, and the cells were washed with RNase-free PBS. Trypsin (200 L) was then added to each well of a six-well plate, and the cells were digested at 37 . The digestion was halted by adding an equal volume of complete medium, followed by centrifugation at 1,000  g for 5 minutes at 4  to collect the cell pellet. The pellet was resuspended in cryopreservation solution, transferred to a programmable cooling box for controlled freezing, and subsequently shipped on dry ice to Novogene for analysis. Finally, the CUT&Tag libraries passing fragment analyzer quality control were then sequenced on Illumina NovaSeq X Plus.