GSM9598154: TissueExTRM_rep1_ATAC; Mus musculus; ATAC-seq
At designated time points (day 49 or day 71 after transfer), circulating ex-TRM cells were isolated from peripheral blood, and tissue TRM and TCIRC cells were sorted from lungs using flow cytometry based on CD45.1/CD45.2, Ly5.1/Ly5.2, and CD69 expression. Sorted cells (3,000–50,000 per population) were pelleted and processed for nuclei isolation using lysis buffer prior to transposition (ATAC-seq) or RNA extraction (RNA-seq). For ATAC-seq, nuclei were subjected to transposition using the Illumina Tn5 transposase (15027865, 15027866) in a 40 µL reaction for 30 min at 37 °C with shaking. Fragmented DNA was purified with a QIAGEN MinElute kit and PCR-amplified for 11–12 cycles. For RNA-seq, total RNA was extracted from sorted cells and converted into cDNA libraries using a strand-specific RNA-seq kit following the manufacturer's instructions. All libraries were sequenced on an Illumina NextSeq 500/550 platform with 50-cycle paired-end reads.