TIM-1 is upregulated on donor Tcon cells, iNKT and Foxp3⁺T cells after HCT. (A-B) Representative flow cytometry plots of TIM-1 staining on iNKT (top), Tcon cells (middle), and Foxp3⁺ T cells (bottom) isolated from the spleen on day 4 after HCT from naive BALB/c mice (A) or BALB/c mice that received TCD BM and B6 Tcon cells after TBI (B). (C) Table showing accumulative data from 3 independent experiments (n = 9/group) from naive BALB/c mice, BALB/c mice treated with TBI alone, and BALB/c mice that in addition to TBI received TCD BM and B6 Tcon cells. The distinction between donor and recipient cells on day 4 post-HCT was determined by flow cytometry based on H-2Kb and H-2Dd surface staining. For statistical analysis between the donor and recipient cells in the mice that received TBI, TCD BM, and B6 Tcon cells, a 2-tailed Student t test was used (mean ± SEM; P ≤ .0001). Gating was based on TIM-1 fluorescence-minus-one control on recipient BALB/c (H-2^d) cells post-HCT. N/A, not applicable; TCR, T-cell receptor; tet, tetramer.