Extended Data Fig. 3: Stages of positive selection in the thymus. | Nature Immunology

Extended Data Fig. 3: Stages of positive selection in the thymus.

From: Reversal of the T cell immune system reveals the molecular basis for T cell lineage fate determination in the thymus

Extended Data Fig. 3

(a) CD4 vs CD8α expression on whole thymocytes (left) and DP thymocytes (right) from B6 and FlipFlop mice. Numbers within profiles indicate frequency of cells in each box (left) and numbers in histograms indicate MFI (right). (n=7/strain, representative of 7 independent experiments). (b) Quantitative analysis of CD4 and CD8α surface expression on thymocytes from B6 and FlipFlop mice, related to Fig. 3c. Numbers indicate MFI. (c) Intracellular calcium mobilization stimulated by TCR/coreceptor signaling of DP FlipFlop thymocytes. Thymocytes were stimulated with anti-TCRβ biotin conjugated antibody either alone (gray bar) or together with anti-CD4 (blue bar) or anti-CD8α (red bar) biotin conjugated antibodies and CD69- DP thymocytes were analyzed. Black arrow indicates crosslinking by avidin (n=3, representative of 3 independent experiments). (d) CD69 vs CCR7 profile of B6 WT thymocytes. Numbers within the profile show frequency of cells in each stage. (e) CD4 vs CD8α profile of WT and FlipFlop thymocytes at Stages 1-5 of development as shown in Fig. 3g. As shown in the profile, DN thymocytes were excluded from the analysis.

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