GSM5034813: NR4w (non recovered); Mus musculus; RNA-Seq
Immediately after learned helplessness, mice were anesthetized, spleens were recovered, and mice were transcardially perfused with PBS. Briefly, the hippocampi were dissected excluding meninges and choroid plexus, passed through a 70 μm cell strainer (BD Bioscience) and the cell suspension was mixed (vol/vol) to obtain a 30% Percoll/R1 medium [RPMI 1640 medium (Corning) supplemented with 1% FBS (Gibco), 100 IU/mL penicillin (Gibco), 100 μg/mL streptomycin (Gibco), 1 x non-essential amino acids (Gibco), 1 μM sodium pyruvate (Gibco), 2.5 μM β-mercaptoethanol (Sigma) and 2 mM L-glutamine (Gibco)]. The cellular suspension was overlaid on 70% Percoll/R1 medium in a centrifuge tube, and centrifuged at 2000 rpm for 20 min without using the brake. The cells at the interface of the 30/70% Percoll gradient were recovered, washed once, resuspended in R10 medium. For cell sorting, cells were stained for 30 min with FITC-conjugated anti-CD45 antibodies (clone 30-F11, ebioscience) and live dead fixable aqua dead cell dye (L34957, Molecular Probes), and sorted on a FACS Aria II, to recover CD45+Live dead- cells. The libraries were prepared using the Chromium Single cell 5' Library & Gel Bead Kit v1.0 and the Chromium Single Cell A Chip kit (10X Genomics) according to the manufacturer's instructions, and run on an Illumina NextSeq 500.
GEO Accession
GSM5034813