GSM4217282: TAK1 mKO macrophage 3; Mus musculus; RNA-Seq
Single-cell suspensions were prepared from mouse brains 3 days after stroke. Mice were transcardially perfused with sterile 0.9% NaCl. The ipsilesional brain hemisphere was harvested, and cell suspensions were prepared using the Neural Tissue Dissociation Kit and gentleMACS Octo Dissociator with Heaters (Miltenyi Biotec). Suspensions were passed through a 70 μm cell strainer, and fractionated on a 30% and 70% Percoll gradient at 500 g for 30 min. Mononuclear cells at the interface were collected. Isolated mononuclear cells were washed, resuspended at 1×10^6 per mL, and stained with fluorophore-conjugated antibodies and the appropriate isotype controls. Fluorescence-activated cell sorting was performed using the FACS Aria I sorter and FACS Diva software (BD Biosciences). Microglia and macrophages were gated as CD11c–Ly6G–CD11b+CD45low and CD11c–Ly6G–CD11b+CD45high, respectively, and sorted for bulk RNA-seq. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM4217282