title: GSM9327854: adult mice, small intestine, ATAC-seq, LTi-1; Mus musculus; ATAC-seq
library_construction_protocol: Fetal liver and fetal thymus were separated under a stereo microscope, and single-cell suspension was generated by robustly pipetting with a 1000-μl pipette. The cell suspension was then filtered through a 100 μm strainer and resuspended in FACS buffer (2% FBS/0.05% NaN3/PBS)　The collected adult intestines were cut into 1 cm pieces and washed with PBS. After opening the intestines along the longitudinal axis, they were incubated in 30mM EDTA-containing PBS at room temperature for 15 minutes and were immediately washed with EDTA-free PBS to remove epithelium. The tissues were then digested by incubating at 37°C in DMEM (Nacalai Tesque) containing 1 mg/mL Collagenase D (SIGMA) and 0.33 mg/mL DNase I (SIGMA) for 30 min. Dissociated cells were collected every 30 min and strained with a 100μm strainer, and then lymphocytes were further enriched on a 40%/80% Percoll gradient. Bone marrow was isolated from the femur and tibia (four bones in total), crushed using the syringe plunger flange, filtered through a 100 µm cell strainer, and resuspended in FACS buffer. After staining by antibodies,  cell sorting were performed with  MA900 (Sony Imaging Products & Solutions). 100-200 cells were used for ATAC-seq following the manufacturer's instructions (Active Motif, #53150).