GSM9155441: p16M10; Mus musculus; RNA-Seq
Both male and female mice were used, 6 mice per group. Briefly, cortical samples were enzymatically digested with the Neural Tissue Dissociation kit (P; 130-092-628, Miltenyi Biotec), and myelin was removed with magnetic bead separation (Myelin Removal Beads II, 130-096-733, Miltenyi Biotec). After myelin removal and red blood cells were removed with Red Blood Cell Lysis Solution (130-094-183, Miltenyi Biotec). Microglia and debris were then removed by magnetic separation using anti-CD11b+ microbeads (130-093-634, Miltenyi Biotec), followed by a dead cell removal kit (130-090-101, Miltenyi Biotec). The final cell pellet was suspended in DPBS supplemented with 0.04% BSA. In all samples, more than 90% of the cells were viable based on Trypan Blue examination. Approximately 6,000 cells from one animal were loaded into each sample well on a chip and combined with gel beads containing barcoded oligonucleotides using a 10x Chromium Controller (10x Genomics). Single-cell libraries were constructed according to the manufacturer's instructions and were sequenced by an Illumina NovaSeq Sequencing System at an average depth of 50,000 reads per cell.