Hs 2100 bioanalyzer

Approximately 50000 viable CD8⁺ T cells sorted from cocultures were used for each library preparation. Cells were lysed in 1 × Lysis Buffer and nuclei were isolated by centrifugation. A TruePrep™ DNA Library Prep Kit V2 for Illumina (Vazyme Biotech) was used to construct transposase‐treated libraries. The mass concentration and molar concentration of the libraries were determined using a Qubit 3.0 Fluorometer and a StepOnePlus™ Real‐Time PCR system, respectively, and the lengths of the inserted fragments were determined using an Agilent HS 2100 Bioanalyzer. Qualified libraries were sequenced using an Illumina HiSeq X ten platform in pair‐end 150 bp style.
Raw data were stored in FastQ format, including the base sequence and corresponding quality information. Trimmomatic (v0.36) was used to remove adaptor‐polluted or low‐quality bases, and reads considered too short (<36 nt) were filtered out to obtain clean data. The clean data were then mapped to the reference mouse genome (mm10) by Bowtie2, and visualized by IGV (Integrative Genomics Viewer). Peaks corresponding to open genomic regions were detected by MACS2, and significantly differential peaks between samples were acquired using MAnorm. The enrichment analysis of the GO term (http://geneontology.org/) was based on a hypergeometric test with a threshold of q < 0.05, to detect significantly enriched pathways.

Approximately 50,000 living cells were used for each library preparation. The cells were lysed in lysis buffer to obtain nuclei, and the TruePrep™DNA Library Prep Kit V2 for Illumina (Vazyme Biotech) was used to construct transposase-treated libraries. The mass concentration and molar concentration of the libraries were determined with a Qubit 3.0 Fluorometer and the StepOnePlus™ Real-Time PCR system, respectively, and the lengths of the inserted fragments were determined with an Agilent HS 2100 Bioanalyzer. Qualified libraries were sequenced on the Illumina HiSeq platform in paired-end 150 bp mode. The data were mapped to the human reference genome (ucsc.hg19) with Bowtie2 (version 2.2.3), and binding sites were identified by using MACS2 (version 2.1.1) with the following parameters ‘-q 0.05 –nomodel –extsize 150 –keep-dup all –call-summits’. ATAC-seq data were shown in Supplementary Data Sheet 2 and the quality control of ATAC-seq was in Figure S2.