Bone marrow Tregs with an immunosuppressive phenotype are mobilized into the blood during SCM. FoxP3-GFP-DTR mice were treated with G-CSF and AMD3100 or PBS for 5 days. BM and blood were collected from the treated mice after mobilization. The number and phenotype of Tregs in the BM, blood, and spleen were assessed by flow cytometry. (A) Total number of CD8 T cells, conventional CD4 T cell, and Tregs in the BM and blood. Tregs were identified using expression of green fluorescent protein (GFP). (B) Representative FACS plots of Tregs in the BM and blood from naïve mice. The proportion of TIGIT⁺Nrp-1⁺ and TIGIT⁺KLRG-1⁺ are shown. (C) Total number of TIGIT⁺Nrp-1⁺ and TIGIT⁺KLRG-1⁺ Tregs in the BM and blood with and without mobilization. (D) Heatmap depicts the expression levels of each marker on Tregs in mobilized and nonmobilized blood. The total number of each population is shown. (E) CD8/Treg and CD8/immunosuppressive Tregs (Pop3) with or without mobilization. Data are combined from 2 independent experiments. Data represents mean ± SEM. Mann-Whitney test for 2 sample comparison. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001.