GSM9435528: B6_P21_01; Mus musculus; RNA-Seq
Tissues were digested enzymatically using Papain and Deoxyribonuclease I (Worthington Biochemical Corporation, Cat. no. LK003153) for 20-30 min at 37°C and stopped using Earl's balanced salt solution (EBSS, Thermo Fisher Scientific, Cat no. 24010-043). Cells were triturated using an 18-gauge needle, centrifuged at 300g at 4 °C, washed with cold DMEM, and filtered using a 100 μm Flowmi Cell Strainer (H13680-0040, Bel-Art. Cells were resuspended in cold DMEM and placed on ice immediately. Single-cell and single-nucleus RNA-seq was performed at the single-cell sequencing core in Columbia University Genome Center. Single cells or nuclei were loaded into chromium microfluidic chips with v3 chemistry and barcoded with a 10x chromium controller (10x Genomics). RNA from the barcoded cells was reverse-transcribed, and sequencing libraries were constructed with a Chromium Single Cell v3 reagent kit (10x Genomics). Sequencing was performed on NovaSeq 6000 (Illumina).