Treg expanded through DR3 signaling show a distinct immunophenotype. Whole T cells were isolated from αDR3 or isotype control Ab-treated WT-B6 mice and analyzed by FACS at 4 days after treatment. (A) The expression level of activation molecules in Foxp3⁺Treg (i), CD4⁺Foxp3^-Tcells (ii), CD8⁺ T cells (iii) after in vivo expansion through DR3 signaling (n = 5; *P < .05; **P < .01; **P < .001; ****P < .0001). (B) The representative results of viSNE analysis using multicolor cytometry data in isotype control Ab-treated Foxp3⁺Treg (Iso-Treg) and αDR3-treated Foxp3⁺ Treg (DR3-Treg). The colors were scaled to the expression level of each molecule as indicated.