Surface molecule expressions on Kaede-red and Kaede-green cells.
(A) Chemokine receptor expression. Skin DLN cells were prepared from the mice sensitized, challenged, and photoconverted as in Figure 3A. These LN cells were stained with isotype-matched control, CCR4, CCR5, and CCR7 mAbs, and the expression levels of Kaede-red and Kaede-green Tregs were evaluated by flow cytometry. (B) Transwell assay. DLN cells were transferred to the upper chamber of the transwell, and CCL17 or CCL21 was added to the lower chamber. The cells were incubated for 3 hours, and the numbers of Kaede-red and Kaede-green cells that migrated to lower chamber were analyzed by flow cytometry. Data are presented as means ± SD and representative of 2 independent experiments. Student’s t test was performed between the indicated groups. *P < 0.05. (C) Surface molecule expression. LN cells were stained with isotype-matched control, CD62L, CD44, CD69, CD25, and CD103 mAbs, and the expression levels were evaluated by flow cytometry. These data are representative of 3 independent experiments.