GSM7804465: glomerulus, 129 strain, Akita-Renin Tg; Mus musculus; RNA-Seq
Mouse kidneys was perfused, excised, minced, digested to obtain single cell suspension as described before. Cells were collected by centrifugation, subjected to live-death dye and sorted by flow cytometry using BD FACS ARIA system. For glomerular samples, mice were perfused with magnetic beads and glomeruli were isolated from kidney tissue by digestion. Then single cell suspension was obtained by further digestion from glomeruli. Isolated living single cell suspensions were converted to barcoded scRNA-seq libraries by using the Chromium Single Cell 3′ Library, Gel Bead & Multiplex Kit, and Chip Kit V3, loading an estimated 7,000–12,000 cells per library/well, and following the manufacturer's instructions. Indexed libraries were sequenced using Illumina NovaSeq 6000 platform, where 150bp pair-end sequences were obtained.