Enhanced MAdCAM-1 and reduced VCAM-1 binding in the absence of TGF-β signaling. ICAM-1 (intercellular adhesion molecule-1) mainly binds to β2-containing integrins and serves as a negative control. MAdCAM-1 binds to α4β7 while VCAM-1 binds to α4β1. Same experimental setup as in Fig. 1. Splenic P14 T cells were incubated with either ICAM-1-Ig, MAdCAM-1-Ig, or VCAM-1-Ig in the presence of GP_33-41 at 37°C for 5 min followed by standard flow cytometry staining and analysis. (A) Representative FACS profiles of pre-gated WT control and Tgfbr2^−/− P14 T cells are shown (three repeats, n = 5/each). (B–E) The percentage of MAdCAM-1 binding (B and D) or VCAM-1 binding (C and E) at day 10 (B and C) and day 23 (D and E) in pre-gated Tcf-1⁺ and Tcf-1⁻ P14 T cells are shown. Each pair of symbols in B–E represents the results from an individual recipient (three repeats, n = 5/each). *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 by paired Student’s t test.