GSM7333221: day 4, control, replicate 2; Mus musculus; RNA-Seq
Four days after stroke onset, brains were removed and sliced in ice-cold artificial cerebrospinal fluid. Peri-infarct area was trimmed and treated with pronase (Sigma-Aldrich) for 75 minutes at room temperature. Enzyme-treated tissue was dissociated by gentle pipetting with a Pasteur pipette. Neuron enriched cell populations (Thy1+/ACSA2-/CD31-/CD45-/O4-/CD140a-/CD140b-) were collected by fluorescence-activated cell sorting (BD FACS Aria III). RNA libraries for scRNA-seq were prepared using Chromium Next GEM Single-cell 3' Reagent Kits v3.1 (10x Genomics) according to the manufacturer's protocol.