Supplementary Figure 4: Detection of Trav11Traj18 transcripts and DN NKT cells in the thymus of Cd4-Cre+Rag2fl/fl mice. | Nature Immunology

Supplementary Figure 4: Detection of Trav11Traj18 transcripts and DN NKT cells in the thymus of Cd4-Cre+Rag2fl/fl mice.

From: Alternative pathway for the development of Vα14+ NKT cells directly from CD4–CD8– thymocytes that bypasses the CD4+CD8+ stage

Supplementary Figure 4

(a) Viable 7-AAD-B220-CD1d dimer-TCRβ-/lo DP and DN thymocyte fractions that were electronically gated as CD5loCD69- cells were sorted from Cd4-Cre+Rag2fl/fl mice, and Trav11Traj18 mRNA expression was determined by qPCR analysis. Relative gene expression (mean ± SEM, n = 3) with Gapdh used as internal control. Results are representative of two experiments, (n.d.) not detected. (b) Detection of CD1d dimer+ DN NKT cells in the thymus, and (c) in the liver from Cd4-Cre+Rag2fl/fl mice, where littermate mice were used as controls. MACS depletion of CD4+, CD8+ and DP cells with magnetic microbeads was done prior to the cell staining. Numbers on flow cytometry plots show the percentage and the absolute number of CD1d dimer+TCRβ+ NKT cells among electronically gated DN cells. Results shown are from one experiment with two mice per genotype analyzed.

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