GSM7662656: S-28,uIRI45min,28d,scRNA-seq; Mus musculus; RNA-Seq
Flow cytometry sorting (FACS)of monocyte or macrophages from kidneys, spleen, and Blood at different time points after uIRI 45 minutes.F4/80+ cells and Cd11b+ cells were sorted separately from kidney samples and mixed at the ratio of 1:1. Cd11b+ cells and Ly6c+ cells were sorted separately from blood samples and collected at a ratio of 1:1. Cd11b+ cells were sorted from spleen samples. FACS-sorted cells were resuspended to a final cell concentration of 700~1200 cells/μl with more than 85% viability as determined by CountessTM II (Thermo Fisher Scientific). 8,000 to 12,000 cells were captured in droplets (10x Genomics Chromium Single Cell V2,V3,3ʹ Reagent Kits). After the reverse transcription step, fifty nanograms of amplified cDNA was fragmented and end-repaired, and sequenced on an Illumina platform using 150 paired-end reads at a coverage of 40,000 mean reads per cell.