GSM6190677
To prepare nuclei, around 50,000 cells at 500g for 5 min, which was followed by a wash using 50 μL of cold 1× PBS and centrifugation at 500g for 5 min. Cells were lysed using cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630). Immediately after lysis, nuclei were spun at 500g for 10 min using a refrigerated centrifuge.  Pools of 5,000-10,000 cortical pyramidal neurons were sorted directly into TRIzol-LS buffer (Invitrogen) and RNA was extracted according to the manufacturer's protocol. 10 ng total RNA was used for library preparation using the SMART-Seq v4 Ultra Low Input RNA Kit (Clontech) and the Nextera XT DNA Library Preparation Kit (Illumina) according to the manufacturer's protocols.