Effects of rATG treatment on CD4⁺CD25⁻ T cells. (A) rATG converted CD4⁺CD25⁻ T cells to CD4⁺CD25⁺ T cells and conferred FOXP3 expression; representative flow cytometric results are shown. (B) Increased FOXP3 and NFAT1 expression in CD4⁺CD25⁻ T cells with rATG detected by Western blotting; results are representative of 3 independent experiments. (C) Proliferation of CD4⁺CD25⁻ T cells promoted with rATG. CFSE intensity was used to evaluate the proliferation. CD3/28 mAb stimulation or rIgG treatment was used as a positive or negative control, respectively. Results are representative of 3 independent experiments. (D) CFSE dilution in rATG-treated CD4⁺CD25⁻ T cells was evaluated based on expression of CD25; results are representative of 3 independent experiments.