GSM8215398: E22-CUS; Mus musculus; RNA-Seq
Brains were collected and processed following the same protocol as used for slice electrophysiology, except for only NMDG aCSF being used in all steps (for details, see section Acute Slice electrophysiology). Tissue corresponding to mPFC was dissected and dissociated into a single cell suspension, as previously described (Munoz, Cell Reports 2017). Tissue was dissociated using the Papain dissociation system (Worthington) following the manufacturer's instructions. Two mice were used for each Sst+ sample to ensure sufficient RNA (~4K sorted neurons per sample), while Camk2+ samples included one mouse each (~40K sorted neurons). Cell suspension obtained was filter (35 μm filter) and kept in ice-cold oxygenated NMDG aCSF, with 0.2% BSA, 0.3% glucose and DNAse. To avoid low quality cells, only fluorescent, but DAPI negative, cells were sorted directly into lysis buffer provided in mRNA extraction kit (RNeasy Plus Micro Kit, Qiagen # 74034). Samples were vortex, immediately processed according to manufacturer's instructions, and eluted RNA was kept in -80°C until sequenced. RNA libraries for RNA-seq were prepared using SMART-Seq v4 Ultra Low Input RNA plus Nextera XT DNA Sample Preparation