GSM9422461: D1 Female CWDS Susceptible 3; Mus musculus; RNA-Seq
24h after social interaction testing, four 14 gauge punches containing NAc were collected from D1-Cre-RT and A2A-Cre-RT mice and snap frozen at -80C.  Polyribosomes were immunoprecipitated as previously described [25], each sample pooled from four mice with similar SI ratios. Homogenates were incubated with 5 μl anti-HA antibody (Biolegend 901515) at 4°C overnight with rotation. Samples were then incubated with 400 uL of protein G magnetic beads (Life Technologies, Carlsbad, CA, USA #100.09D) overnight at 4°C with rotation. Beads were washed in a magnetic rack with high-salt buffer. RNA was extracted with a DNase step (Qiagen, Germantown, MD, USA) using the RNeasy mini kit (qiagen) by adding RLT and following manufacturer instructions.  RNA concentration and quality were assessed via Bioanalyzer (Agilent). Samples with RNA integrity number >8 were used and pooled into biological quintuplicates for sequencing at the Maryland Genomics core of the UMSOM Institute for Genome Sciences (IGS) and processed as described previously [26]. Libraries were prepared from 10 ng of RNA from each sample using the SMART-Seq v4 kit (Takara)