GSM9291773: CeA_snRNAseq_Sensitive_M7; Rattus norvegicus; RNA-Seq
Rats were immediately sacrificed after last footshock session, and brains were fresh frozen and then stored at −80°C until further processing. We sectioned the brains at a cryostat (Leica CM3050s) and collected 300 µm slices from approximately bregma -1.6mm to -2.8mm based on the rat brain in stereotaxic coordinates. Then CeA tissue was harvested bilaterally from the brain slices using a 1.5 mm diameter biopsy punch and tissue blocks were stored at −80°C. Nuclei were isolated using the Chromium Nuclei Isolation Kit with RNase inhibitor (10X Genomics) following the manufacturer's instructions with a lysis time of 6.5 minutes and a single wash following debris removal. The nuclei were resuspended in a final volume of 50–100 µL at a target concentration of 500–1,200 nuclei/µL. Single-nucleus RNA-seq libraries were generated using the Chromium Next GEM Single Cell 3′ Kit v3.1 (10x Genomics), targeting 10,000 nuclei per sample. Libraries were sequenced on an  Illumina NovaSeq 6000 system using an S1 flow cell, producing approximately 500 million reads per sample.