Single-cell RNA transcriptomics of thymic iNKT cells from BALB/c and Cd80/Cd86 -/- (B7KO) mice

Single-cell RNA-seq analysis of murine thymic iNKT cells from three independent BALB/c mice and three independent Cd80/Cd86 (B7)-deficient mice Processed thymocytes were enriched for PBS57-CD1d reactive cells by incubating thymocyte cell suspensions with PE conjugated PBS57-CD1d tetramers for 45 minutes at 4C, then incubated with anti-PE Ab and magnetic microbeads (STEMCELL) for 15 and 10 minutes at RT, respectively, followed by separation by using an EasySep magnet (STEMCELL) according to manufacturer’s instructions. A 50:50 mix of CD44low and CD44high PBS57-CD1d tetramer+ cells were sorted using a BD FACS Aria. Single-cell RNA-seq was performed using 10X Genomics Chromium isingle-cell controller and 5' gene expression kit.