Quantitation of FoxP3 expression in donor cells infiltrating lymphoid tissues and target organs on days 10, 13, and 16 after transplantation. (A) Transplants consisted of WT TCD-BM and WT CD25^- T cells, with column-purified WT/eGFP (▪) or CCR5^-/-/eGFP (▨) T_regs. The numbers of eGFP⁺CD4⁺ cells infiltrating the spleen and liver were determined by flow cytometry on day 10. These data are representative of 2 similar experiments. ^*P < .05 (B) FoxP3 expression in donor cell infiltrates purified on day 10 from spleens and livers of transplant recipients receiving TCD-BM and WT CD25^- T cells, either alone (▦) or with column-purified T_regs from WT/eGFP (▪) or CCR5^-/-/eGFP (▨) donors. Donor cell infiltrates were pooled from 3 mice per group. Data shown are representative of 2 separate experiments yielding similar results. ^*P < .05. (C) FoxP3 expression in donor cell infiltrates purified on day 10 from lungs of mice transplanted as above. Donor cell infiltrates were pooled from 3 mice per group. (D) FoxP3 expression in donor cell infiltrates purified on days 13 and 16 from spleens, livers, and MLNs of recipient mice transplanted as in panel B. Donor cell infiltrates were pooled from 4 mice per group on day 13 and from 3 mice receiving WT/eGFP T_regs and 2 mice receiving CCR5^-/-/eGFP T_regs on day 16. (E) Mice underwent transplantation as in panel A, and intracellular Scurfin analysis was performed on leukocytes pooled from the spleen or liver of 3 mice per group on day 10. Total GFP⁺CD4⁺Scurfin⁺ cells per organ were calculated by multiplying the total number of GFP⁺CD4⁺ cells in each organ, by the percent Scurfin⁺. Data shown are mean ± SEM. ^*P < .05.