GSM8417118: RPTEC, TET2 knockdown, RNAseq; Homo sapiens; RNA-Seq
For RNA seq: RNA was extracted using Rneasy mini kit (Qiagen); for snRNAseq, Nuclei were extracted from flash-frozen kidney tissue. Briefly, 10-30 mg frozen kidney tissue were minced on ice into pieces in lysis buffer (Tris-HC, NaCl, MgCl2, NP40, and RNAse inhibitor), transferred into a gentleMACS C tube and homogenized using gentleMACS homogenizer. Next, the homogenized tissue was filtered through a 40 μm strainer (Fisher Scientific, #08-771-1) and then centrifuged at 500 × g for 5 mins at 4°C. The pellet was resuspended in wash buffer (BSA, RNAse inhibitor, 1× PBS) and then filtered through a 40 μm Flowmi cell strainer (Sigma Aldrich, #BAH136800040-50EA). RNA-Seq libraries were generated using the NEB's non-directional RNA seq kit; For snRNAseq libraries, the cDNA libraries were constructed using the 10X Chromium library construction kit (PN-1000190).