Fig. 2.

Decrease of the KLRG1⁺ CD103⁺ subpopulation, IL-10 production, and CTLA-4 expression in Ti-Treg cells by Met administration. (A) Tumor growth in mice inoculated with MethA, which were given Met (blue circles) or normal feed (red circles) as indicated by the shadowed rectangle. N = 10 for each group. (B) Terminally differentiated, a KLRG1⁺ CD103⁺ population in Ti-Treg cells was reduced by Met administration. Treg subpopulations in the spleen, tumor-draining lymph node (TdLN), and TILs of the MethA-bearing mice were identified by their expressions of KLRG1 and CD103. The experiment was performed twice, and the flow cytometry panels are representative of five independent mice in one experiment. Bar graphs indicate % ratios of the KLRG1⁺ CD103⁺ population among CD4⁺ CD25⁺ Foxp3⁺ Treg cells in one experiment. N = 5 for each group. (C) IL-10 production and cell surface CTLA-4 expression were reduced in Ti-Treg cells but not in Treg cells from the spleen, TdLN by Met administration. The cells were pooled from three mice for each group and analyzed by flow cytometry. Mean fluorescence of IL-10 and CTLA4 is shown as bar graphs as indicated. N = 3 for each group. The results are representative of three independent experiments. +, Met administered; −, Met not administered; *P < 0.05; **P < 0.01.