Effect of Dec on FOXP3 expression in anti-CD3/CD28 antibody coated bead-activated CD4⁺CD25⁻ T cells. (A) Both human and mouse CD4⁺CD25⁻ T cells express FOXP3 after Dec treatment in the presence of anti-CD3/CD28 antibody coated beads. (B) Real-time RT-PCR for human FOXP3 mRNA was performed in triplicate at various times after Dec treatment of activated T cells. Levels of mRNA for FOXP3 increase each day and are comparable to that seen in bead-activated Tregs (brown) by day 4. (C) CD4⁺CD25⁻ T cells obtained from Foxp3-ires-GFP KI mice confirm induction of FOXP3 expression after Dec and AzaC treatment (indirectly measured as GFP expression). (D) DcT FOXP3 expression (blue) persists for at least 7 days, suggesting prolonged and stable expression of FOXP3 in vitro. Treg (red) is shown for comparison. A pool of 2 independent experiments.