Nuclei were isolated from fresh frozen human occipitotemporal cortex brain tissue using a sucrose gradient ultracentrifugation protocol. Samples were a subset of those originally reported in (Gerrits et al., Acta Neuropathologica, 2021). Nuclei were separated into 3 populations using FANS: NeuN+ nuclei, Olig2+ nuclei, and the double negative nuclei. We collected 30,000 NEUN+ nuclei, 15,000 OLIG2+ nuclei, and 5,000 double negative nuclei. All collected nuclei were then loaded onto the Chromium v3 single cell chips (10x Genomics). This strategy has previously been used to enrich for less abundant CNS cell types (Gerrits et al., Acta Neuropathologica, 2021). Single-nucleus cDNA libraries were constructed and dual-indexed using the Chromium Single Cell 3′ Reagents Kit v3 and corresponding user guide (10× Genomics).