Foxp3⁺ Tregs were not involved in the inhibitory function of DCapos both in vitro and in vivo.

<p>(A) Left, DO11.10 CD4⁺ T cells (2×10⁵ cells) with Foxp3^EGFP were purified, and the CD4⁺CD25⁺Foxp3⁺ cells were measured in a FACS system (numbers indicate percentages). These cells were then co-cultured with OVA_323–339-loaded mDCs (2×10⁴) in the presence or absence of DCapos (2×10⁴ cells) for 5 days in vitro. On the fifth day, the cells were harvested and the numbers of CD4- and KJ1-26-positive cells were determined by FACS analysis (black). The proportion of CD4⁺KJ1-26⁺CD25⁺Foxp3⁺ cells was determined by FACS (grey). Data are presented as mean ± SD of triplicate wells, and the results represent 2 separate experiments. (B, C) Apoptotic cells (AC), DCapos, or phosphate-buffered saline (PBS) was injected into the peritoneal cavity of Foxp3^EGFP mice (PBS, <i>n = </i>5, AC, <i>n = </i>8, DCapo, <i>n = </i>5). The CD4⁺CD25⁺Foxp3⁺ cells in the peripheral blood were assayed by FACS at days 0, 1, 4, and 11 (numbers indicate percentages). Data in b–c are representative of 3 separate experiments. *<i>p</i> > 0.05.