GSM9831159: Kidney tubulointerstitium, Nphs2 R231Q/A286V, male, age 7\5; Mus musculus; RNA-Seq
Kidneys were extracted, perfused with magnetic beads (ThermoFisher), minced and enzymatically digested. The suspension was filtered and washed, and glomerular and tubulointerstitial fractions were separated magnetically. Nuclei were isolated from the tubulointerstitial fraction using Nuclei EZ Prep (Sigma, NUC101-1KT): cells were lysed in EZ lysis buffer supplemented with 1× protease inhibitor mix (Roche) and 0.1% RNase inhibitor (NEB, M0314L). Single-nucleus suspensions were loaded on the 10x Genomics Chromium controller and gene-expression libraries were prepared with the Chromium Single Cell 3′ Gene Expression v3.1 (Dual Index) kit per the manufacturer's instructions.