(A–E) The DLN cells of Kaede/Foxp3hCD2/hCD52 mice photoconverted on the abdomen 24 hours prior were stained with CD4, CD25, and hCD2 mAbs. Shown here are the flow cytometric plots for hCD2/Foxp3 and CD25 staining among CD4+ cells (A) and Kaede-red and Kaede-green expression on hCD2+CD4+ cells among skin DLN cells (B). (C) The DLNs and non-DLNs from the mice 24 hours after photoconversion were stained with CD4, hCD2, and CD44 mAbs and subjected to flow cytometry. (D) hCD2/Foxp3 expression in total (Kaede-red plus Kaede-green), Kaede-red, and Kaede-green CD4 cells was compared by flow cytometry. (E) The numbers of CD44mid naive (M), CD44hi memory (H), and naive plus memory (H/M) phenotypes of hCD2–CD4+ non-Tregs (–), hCD2+CD4+ Tregs (+), and total (hCD2– and hCD2+; +/–) CD4+ T cells in total CD4+ (Kaede-red plus Kaede-green) cells and Kaede-red cells in the DLNs were counted. Data are presented as means ± SD and are representative of 3 independent experiments. Student’s t test was performed between the indicated groups. *P < 0.05. Numbers within plots or histograms indicate percentage of cells in the respective areas (A–D).