Figure 4. Antibody-based FACS and RNA-Seq for CNS cell types from adult brain tissue.

(a) Left panel shows forward/side scatter (FSC/SSC) of dissociated cortical tissue. DAPI⁺ events (‘cells') were gated to select for nuclei-containing singlets (second panel), and neurons and microglia were isolated as two distinct populations corresponding to NeuN⁺CD11b^– and NeuN⁻CD11b⁺ cells, respectively (third panel). The NeuN⁻ population was further used to isolate GFAP⁺ astrocytes (last panel). (b) RNA-Seq data confirmed that NeuN⁺ sorting enriched for cells expressing neuronal markers, GFAP⁺ sorting enriched for cells expressing astrocytic markers, and CD11b⁺ sorting enriched for cells expressing microglial markers (n=5 animals; one astrocyte sample was excluded from the analysis due to evidence of neuronal contamination). Bars represent mean±s.d. (Prism).