GSM8068290: brain, Plexin-B2_cKO, rep1, scRNAseq; Mus musculus; RNA-Seq
Plexin-B2 knockout was induced by administrating Tamoxifen (100 mg/kg) 3 days before intracranial injection of KR158 GBM cells and every other day until 8 weeks. After 8 weeks, mice were euthanized and the brain tissues containing KR158 tumor were harvested on ice. Brain tissue pieces were taken from approximately 1 mm anterior to 1 mm posterior from the injection site and minced with scalpel blades and dissociated into single cell with papain digestion. Debris from the brain tissues were removed by Percoll centrifugation and red blood cells (RBCs) were lysed with RBC lysis solution on ice for 10 minutes. Cells were washed and resuspended in FACS buffer containing 0.2% BSA and 20 μg/ml DNase and submitted for single cell sequencing with the 10X Genomics system at the Mount Sinai Genomics core facility. GEM formation, cDNA amplification and library preparation was performed according to the manufacture's instructions (Chromium Next GEM Single Cell 3ʹ Kit v3.1, 16 rxns PN-1000268; Chromium Next GEM Chip G Single Cell Kit, 48 rxns PN-1000120). Barcoding libraries was performed per User guide, CG000315 from 10x Genomics (Dual Index Kit TT Set A, 96 rxns PN-1000215).