brain-mouse1
Brains were dissected from perfused mice, minced, and enzymatically digested (papain, 37C, 20 min) with or without transcriptional inhibitors (Actinomycin D, Triptolide, Anisomycin). Cells were dissociated, filtered (70 m), and myelin debris was removed via Percoll gradient centrifugation. Live cells were enriched using Miltenyi Dead Cell Removal Microbeads and resuspended in PBS/0.2% BSA. Single-cell 5 libraries were prepared (10x Genomics Chromium Single Cell 5 v2) and sequenced on an Illumina NovaSeq X Plus.