GSM8784949: Periarticular tissue Ctrl 2; Mus musculus; RNA-Seq
The periarticular tissue was minced and digested in RPMI 1640 medium (Gibco) supplemented with 1.5 mg/mL collagenase IV (Sigma), 0.5 mg/mL DNase I (Roche), and 2% FBS for 90 min at 37°C with gentle shaking.To obtain sufficient immune cells for analysis, four mice were pooled as one sample for the control group (12 control mice in total for n=3), and then CD45+ and CD45- cells were sorted and mixed at a ratio of 3:1 in both groups for scRNA-seq analysis. The cells were loaded on a 10X Chromium Controller (10X Genomics) according to the manufacturer's protocol to generate Gel-Bead in Emulsions (GEMs), which were used to generate barcoded, full-length cDNA through reverse transcription reactions. Barcoded scRNA-seq libraries were prepared using the Chromium Single Cell 3′ Reagent Kit. After final preparation and QC performed, the libraries were sequenced on a NovaSeq 6000 platform (Illumina), and 150 base pair (bp) paired-end reads were generated.