GSM4551971: Sst_Kir2.1WT_rep2; Mus musculus; RNA-Seq
Sst-cre; Sun1 or Vip-cre; Sun1 mouse cortices were harvested and fixed in RNAlater overnight at 4°C. Cortices were homogenized with a dounce-homogenizer and tight pestle 15 times in 5 mL Buffer HB (0.25 M sucrose, 25 mM KCl, 5 mM MgCl2, 20 mM Tricine-KOH pH7.8, 1mM DTT, 0.15 mM spermine, 0.5 mM spermidine, Roche protease inhibitor tablets, 10mM Sodium Butyrate, 1 mM Rnase Inhibitor when doing downstream RNA-seq). IGEPAL CA-630 (Sigma) was added to a final concentration of 0.3% to the homogenate, followed by douncing five more times with tight pestle. Homogenized cortices were filtered through a 40 µm strainer, and centrifuged at 500g to pellet the nuclei. The nuclei were resuspended in FACS buffer (1XPBS, 1% BSA) and sorted on GFP-positive nuclei (using Sun1-GFP localizes throughout the nuclear membrane) were sorted into cold FACS buffer using a Sony SH800Z Cell sorter. All single nuclei RNA sequencing experiments were performed using the Chromium TM Single Cell 3' Library kit v2 (10X Genomics) following manufacturer instructions.
GEO Accession
GSM4551971