GSM8565270: snRNAseq_d30pi_combined; Mus musculus; RNA-Seq
Female CD45.1+ CD8+ P14 T cells were adoptively transferred into male CD45.2+ recipients (1×105 cells/mouse) 30 minutes prior to infection with LCMV-Armstrong. At 28 days post infection, mice were euthanized and small intestine was dissected, flayed, and washed in cold PBS; Peyer's patches were excised from the small intestine. The small intestine was divided into three equal sections designated as the proximal, middle, and distal small intestine. Tissue sections were cut into ~3 mm pieces and flash-frozen in liquid nitrogen for 2 min. Nuclei isolation was performed with 10x Genomics Chromium Nuclei Isolation Kit per the manufacturer's instructions. Briefly, 30-50 mg of flash-frozen tissue per sample was dissociated with a pestle, incubated for 10 minutes on ice, and washed. Dissociated tissue was passed through a Nuclei Isolation Column, and flowthrough nuclei were washed in Debris Removal Buffer and Wash and Resuspension Buffer. Nuclei were quantified with Nexcelom Bioscience Cellometer. For maximum targeted recovery, 40,000 nuclei per sample were loaded for GEM generation. Samples were processed via the Chromium Next Gem Single Cell 3' HT Dual Index v3.1 protocol and sequenced to a depth of 550 million read pairs per sample (~23,000 read pairs per nucleus) on a NovaSeq 6000 system (Illumina).