RNAseq analysis of thymic CD4-CD8- double-negative (DN) invariant NKT cells from CD4+CD8+ double-positive (DP)-specific Rag2-conditional knock-out mice

We report the identification of DN NKT cells developed from DN stage thymocytes. We analyzed the gene expression profiles of NKT cells that have developed from DN stage thymocytes isolated from DP-specific E8III-Cre transgenic Rag2-floxed mouse strain, and NKT cells developed from DP thymocytes sorted as YFP-reporter positive NKT cells that were isolated from E8III-Cre transgenic Rosa26-loxP-STOP-loxP mouse. Overall design: Next generation sequencing of total transcriptomes using total RNA isolated from FACS sorted ex vivo thymic and liver NKT cells. DN-origin NKT cells were obtained from DP-specific E8III-Cre transgenic Rag2-floxed mice, and DP-origin NKT cells were obtained from E8III-Cre transgenic Rosa26-loxP-STOP-loxP mouse. The sorting gate was determined as: 7AAD-CD45R-CD4-CD8-CD1d dimer+TCRb+ cells.