Fig. 3

SENP3 is required for the suppressive function of Treg cells. a CFSE-labeled wild-type CD4⁺CD25^–YFP^– T cells were cultured with the indicated number of Senp3^+/+Foxp3-Cre or Senp3^fl/flFoxp3-Cre Treg cells and stimulated with anti-CD3 and anti-CD28 antibodies for three days. Summary graphs of the percentage of proliferating CFSE-labeled T cells. b–d Disease phenotype of 6-week-old RAG-1-deficient mice given adoptive transfer of wild-type naive CD45RB^hi CD4⁺ T cells together with Senp3^+/+Foxp3-Cre or Senp3^fl/flFoxp3-Cre Treg cells. Body weight was presented relative to initial weight (b). Frequency of cytokine-producing CD4⁺ T cells in the mesenteric lymph nodes (c) or hematoxylin-eosin staining of colon sections (d) from recipient mice at 12 weeks after adoptive transfer. Bars, 500 μm. e Tumor growth in Senp3^+/+Foxp3-Cre or Senp3^fl/flFoxp3-Cre mice subcutaneously (s.c.) injected with B16-F10 melanoma cells (n = 8). f, g Flow cytometric analysis of the frequency of IFN-γ-producing CD4⁺ and CD8⁺ T cells or Foxp3⁺CD4⁺ T cells in tumors of Senp3^+/+Foxp3-Cre and Senp3^fl/flFoxp3-Cre mice (day 14 after B16-F10 tumor injection). h Tumor growth in Senp3^+/+Foxp3-Cre or Senp3^fl/flFoxp3-Cre mice subcutaneously (s.c.) injected with MC38 colon cancer cells (n = 8). i, j Flow cytometric analysis of the frequency of IFN-γ-producing CD4⁺ and CD8⁺ T cells or Foxp3⁺CD4⁺ T cells in tumors of Senp3^+/+Foxp3-Cre and Senp3^fl/flFoxp3-Cre mice s.c. injected with MC38 colon cancer cells (day 14). Data are representative of at least three independent experiments. Error bars are the mean ± SEM values. n = 5 or 8. Two-tailed unpaired Student’s t tests were performed. *P < 0.05; **P < 0.01