HC4
RRBS library preparation was performed using the Acegen Rapid RRBS Library Prep Kit (Acegen, Cat. No. AG0422) following the manufacturer's protocol. The DNA samples initially underwent digestion using the methylation-insensitive restriction enzyme; then the fragmented DNA underwent a series of processes including end-repaired, A-tailed, and ligation to sequencing junctions where all cytosines were methylated; following this, fragments of DNA ranging from 150 to 300 base pairs were chosen for cleavage.