GSM9764930: p21_3w_1; Mus musculus; RNA-Seq
For single-cell RNA sequencing (scRNA-seq) analysis, 3-week-old p21 reporter mice were used. p21 reporter mice received a single intraperitoneal injection of tamoxifen (1 mg/mouse) 48 hours prior to analysis. The diaphysis was trimmed approximately 2 mm distal to the growth plate, and metaphyseal regions of distal femurs and proximal tibias were collected in digestion buffer. Following bone dissociation as described above, FACS was performed using the FACS Melody system (BD Biosciences). CD31/CD45/TER119-negative cells were sorted after dead cells were excluded using 7-AAD (1:200; BioLegend, 420403). Sorted cells were immediately collected in ice-cold cell suspension buffer and loaded onto a Chromium Controller (10X Genomics). Single-cell libraries were prepared using the Chromium Next GEM Single Cell 3′ Reagent Kits v3.1 (10X Genomics).