Figure 4: Enhanced capacity of rapamycin-treated H1-DC to promote Treg induction. (a) The starting population of naive CD4+ T cells was analysed by flow cytometry for the expression of CD25 and Foxp3, markers associated with commitment of T cells to the regulatory T cell lineage. (b) Rapamycin enhances the capacity of H1-DC to induce Treg cells compared to moDC. DCs were either untreated, matured with the maturation cocktail or treated with rapamycin for 3 days prior to maturation. DCs were harvested, washed, and plated at 2 × 105 per well with 106 naive CD4+ T cells per well of a 24-well plate to yield a ratio of DC : T cells of 1 : 5. On day 7, cocultures were stained for CD4, CD25, and Foxp3 and analysed. Dead cells were excluded from the analysis using 7-AAD staining. Data from one experiment representative of 4 independent experiments are shown.