Figure 2

Delayed FoxP3 expression in CD4⁺CD8^-CD25^+/- cells in neonatal thymus. A: Generation and characterization of the anti-FoxP3 antibody.293T cells were transfected with either vector or pHSPG-mFoxp3. FoxP3 was specifically detected by Western blot with the anti-FoxP3 antibody (top left), immunofluorescence (IF) staining (lower left), by FACS analysis (middle). The anti-FoxP3 antibody specifically stained intracellular FoxP3 in CD4⁺CD8^-CD25⁺ Treg cells from wild type, but not scurfy, mice (right). B: Expression of FoxP3 protein is not detected in CD4⁺CD8^-CD25⁺ thymocytes until 3 days after birth. Percentages of CD4⁺CD8^-CD25⁺ and CD4⁺CD8^-CD25^- thymocytes (gate on the CD4⁺CD8^- population from Fig. 1A) that expressed FoxP3 are shown, with the control IgG as background.