Fig. 1. Development of Foxp3+ T cells inFoxp3-GFP-Cre × R26-YFPtransgenic mice.
(A)Thymocytes were isolated, stained for CD4 and CD8, and GFP and YFP expression was analyzed by flow cytometry. Thymocytes were gated as CD4+CD8+ double positive (DP) and individual CD4 or CD8 single positive (SP) populations. (B) The histograms show CD24 expression on the CD4SP GFP and YFP populations gated as shown in panel A. (C) GFP and YFP expression by gated CD4+ T cells from lymph nodes (LN) and spleen. (D) Methylation analysis of the Foxp3 locus. The methylation status of the CpG motifs of the TSDR of sub-cloned from purified Tconv, Tregs and exFoxp3 cells. Data is the average of data generated in3 independent experiments. (E) Graphs show the indicated GFP and YFP populations as a percentage of CD4+ T cells in spleen, LN, liver and Peyer’s patch of Foxp3-GFP-CrexR26-YFP mice. Each point represents an individual mouse. Experiments were performed with 6 -9 week old mice and are representative of 4 (A, B) and 20 (C) independent experiments.