GSM7928392: OT-1 cells, wildtype, NOH (GEX); Mus musculus; RNA-Seq
Single-cell suspensions of spleen and lymph nodes of 2 wt and 2 NOH mice at day 7 after infection were pre-enriched for CD8+ cells applying negative selection immunomagnetic cell separation. CD8α+CD90.1+DAPI- OT-1 cells were isolated from pre-enriched cell suspensions by flow cytometry-assisted cell sorting using a FACSAria III cytometer. Sorted OT-1 cells of each sample were labelled with different oligonucleotide-tagged TotalSeq anti-CD45/anti-MHC-I hashing antibodies. Hashed cells were multiplexed and loaded on a GemCode Single-Cell Instrument (10x Genomics, Pleasanton, CA, USA) to generate single-cell gel beads in emulsion (GEMs). Gene expression and feature barcode libraries were generated as described in the User's Guide for 10x Chromium Single Cell Reagent Kits with Feature Barcoding technology for Cell Surface Protein. The library pool was commercially sequenced on an Illumina NovaSeq6000 platform (PE150bp) at Novogene Europe (Cambridge, UK). scRNA-seq (10X platform)