T-cell activation and Th1 bias in recipients of SMAD3-KO grafts. (A) Cytokine bead array analysis of colon extracts on day 40 after AHCT (n = 5). (B-C) Percentage of T-bet⁺ CD4 T cells among live cells on day 40. (B) One representative contour plot. (C) Mean percentage and SEM for 5 mice per group. (D) Mean percentage of CD43i⁺ CD4⁺ T cells among live cells from colon extracts on day 40 after AHCT (n = 7). (E) One representative contour plot showing costaining for CD4 and T-bet on day 20. (F) Percentage of CD4⁺ cells and of CD4⁺T-bet⁺ cells among live cells extracted from the colon on day 20 (n = 8). (G) Percentage of colon CD4 T cells expressing FOXP3 on day 20 and 40 after AHCT. Statistical analyses were performed with Student t test: *P < .05, **P < .01.