We isolated nuclei isolated as previously described.71 Briefly, we homogenized tissues using a loose glass dounce homogenizer followed by a tight glass homogenizer in EZ PREP buffer (Millipore Sigma, Cat# NUC-101). We washed nuclei once with EZ PREP buffer and once with Nuclei Suspension Buffer (NSB; consisting of 1× PBS, 0.01% BSA and 0.1% RNase inhibitor (Clontech, Cat# 2313A)). We re-suspended the washed nuclei in NSB and filtered them through a 35-μm cell strainer (Corning, Cat# 352235). We counted the nuclei and diluted down to 1000 cells/μl. We loaded approximately 10,000 cells from each brain region onto a 10X chip which were then run through a 10x Genomics Chromium controller.
