CD4T_ATAC_Wwp2_Itch_dKO (CRX1855452)
ATAC-seq assay was performed using the Hyperactive ATAC-Seq Library Prep Kit for Illumina (Vazyme Biotech, #TD711) according to the manufacturer's instructions. Briefly, around 1x10^5 living cells were sorted and collected by centrifugation at 500 x g at 4 deg C for 5 min. Cells were resuspended in lysis buffer and incubated for 5 min on ice. Subsequently, the nuclei were pelleted by centrifugation at 4 deg C for 5 min. Pellets were resuspended in transposome mix and incubated at 37 deg C for 30 min for tagmentation. Then, a stop buffer was added to the mixture at 55 deg C for 5 min to stop the tagmentation. Next, tagmentated DNA were bound and selected by magnetic beads, then this magnetic beads were gently rinsed twice with 80% ethanol and tagmentated DNA were eluted with 20ul double-distilled water. Libraries were constructed using NEBNext(R) Ultra(TM) DNA Library Prep Kit for Illumina(R) (NEB #E7370S). And all ATAC libraries were sequenced by KAITAI-Bio using the Illumina NovaSeq 6000 platform in PE150 mode (KaiTaibio, Hangzhou, China).