Figure 3.

RelB^DCko mice do not promote enhanced induction of iTregs. (A) Scheme of experimental setup. Micro-osmotic pumps loaded with PBS or OVA peptide were subcutaneously implanted in RelB^DCko and control mice. One day later mice received an intravenous injection of 4.5–5 × 10⁶ carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled CD4⁺ CD25⁻ OT-II × Rag1^−/− T cells. Twelve days after adoptive transfer, cell suspensions from peripheral LN (PLN) and SPL of RelB^DCko and control mice were stained for CD4 and Foxp3 and analyzed by flow cytometry. (B) Representative dot plots showing CFSE and Foxp3 expressing among transferred CFSE⁺ CD4⁺ T cells in PLN and SPL. Numbers indicate percentages of cells in each quadrant. (C) Percentages of Foxp3⁺ iTregs among CFSE⁺ CD4⁺ T cells in PLN and SPL of RelB^DCko (n ≥ 6) and control mice (n ≥ 5) are depicted. (D) Frequencies of proliferating CFSE^low Foxp3⁻ conventional T cells among CFSE⁺ CD4⁺ Foxp3⁻ T cells in PLN and SPL of RelB^DCko (n ≥ 6) and control mice (n ≥ 5). (B–D) Data represent the mean values + SD from three independent experiments with n ≥ 2 mice per genotype and condition. Statistical analyses were performed using Mann–Whitney test: ns, not significant, p < 0.05.