GSM7437972: cd45_neg_6wk; Mus musculus; RNA-Seq
Joint tissue was digested for 1 hour at 37 ºC in RPMI containing 0.2 mg/ml liberase TL (Roche) and 1 unit/ml DNase (Thermo), inverting every 15 minutes. Cells were then filtered through a 40 µm filter and washed in PBS containing 2% FBS and 1 mM CaCl2. Live cells were obtained through magnetic sorting using the EasySep Dead Cell Removal (Annexin V) Kit (StemCell Technologies) per manufacturer's instructions. Following dead cell removal, cells were washed in MACS buffer (PBS containing 0.5% BSA and 2 mM EDTA) and CD45+ and CD45- cells separated using CD45 MicroBeads and a MiniMACS Separator per manufacturer's instructions (Miltenyi Biotec). RNA libraries were prepared for sequencing using standard Illumina protocols