title: GSM2876262: iR2ko_334; Mus musculus; RNA-Seq
library_construction_protocol: 1. For RNA-sequencing in whole kidneys, total RNA was extracted from whole kidneys from well-perfused mice using an RNeasy mini kit (Qiagen). 2. For sorted macrophages, kidneys from 7-9 m old mice were first minced and digested with collagenase B (Roche) for 45 minutes at 37ºC. Macrophages were enriched by centrifuging cells in 40% percoll/PBS (v/v). Cells were stained with fluorochrome-conjugated anti-mouse antibodies, and CD45+F4/80hiCD11b+Ly6G-Ly6C- kidney macrophages were sorted into RLT lysis buffer directly. Total RNA was extracted using an RNeasy mini kit (Qiagen). 1. For total kidney, library preparation and sequencing were performed by Genomics Resources Core Facility at Weill Cornell Medical College using TruSeq RNA library prep kit (Illumina) followed by single read sequencing on the Illumina HiSeq 2500 instrument (4-6 samples/SR lane, 50 bp reads). 2. For sorted macrophages, library preparation and sequencing were performed at Epigenetics Core facility at Weill Cornell Medical College using the SMART-Seq v4 Ultra Low Input RNA Kit (Clontech) followed by single read clustering and 51 cycles sequencing on the Illumina HiSeq 2500 instrument.