GSM8554632: CD45+ cells isolated from muscle of mice with TB mice; Mus musculus; RNA-Seq
Freshly isolated tissues were finely minced and enzymatically digested with RPMI1640 containing 0.3 mg/ml Liberase TL (5401119001, Sigma-Aldrich, St. Louis, MA, USA) and 0.4 μg/ml deoxyribonuclease I (DN25, Sigma-Aldrich) for 2 h at 37°C with gentle agitation. The digested cells were further disaggregated using 18G and 21G syringes (Korea Vaccine Co., Ltd, Seoul, Korea) and filtration with a 100-μm cell strainer (93100, SPL Life Sciences, CITY, Korea). After red blood cell depletion with ammonium-chloride-potassium lysis buffer (A10492-01, Gibco), CD45+ immune cells were positively isolated with magnetic beads (130-052-301, Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instruction, and then subjected to flow cytometry or scRNA-seq. Single-cell RNA sequencing (scRNA-seq) data were generated by GENINUS Inc. (Seoul, Korea). Barcoded sequencing libraries were prepared using the Chromium Single Cell 5' v2 Reagent Kits (10x Genomics), and sequencing was performed on the Illumina NovaSeq 6000 platform.