GSM9059660: CD45+-positive cells, UNx, 1week; Mus musculus; RNA-Seq
The right kidney was then harvested from each mouse,  and digested by incubation in collagenase D (5 mg/mL; Sigma-Aldrich) for 30 minutes at 37 °C. The digested tissue was  filtered through a 70-µm strainer (Corning), then centrifuged at room temperature (3000 RPM for 30 minutes, no acceleration, no brake) in a Percoll density gradient. After isolation, approximately 1 x 10^6 KMNCs per mouse were pooled within their experimental groups for CD45+ cell sorting and downstream scRNA-seq. Flow-sorted CD45+ kidney cells were partitioned and barcoded using the 10X Chromium Controller, with approximately 10,000 cells loaded onto the NovaSeq 6000 S4 200-cycle flow cell. The library was constructed using the Chromium Next GEM Single Cell 5' HT v2 Kit and sequenced on the Illumina NovaSeq 6000 system.