FIG. 3.
Expression of CD25 and Forkhead box P3 (FoxP3) from naive CD4+ T cells following coculture with ASCs under ambient and low O2 culture conditions. IFN-γ-treated ASCs were cocultured with naive CD4+ T cells for 3 days under low or ambient O2 culture conditions at ratios of 1:100 (1,000 ASCs:100,000 T cells), 1:10 (10,000 ASCs:100,000 T cells), and 1:1 (100,000 ASCs:100,000 T cells). T cells were collected and stained for fluorescein isothiocyanate (FITC)-CD4, phycoerythrin (PE)-CD25, and Alexa Fluor 647-FoxP3 (AF-FoxP3) and flow cytometry was used to calculate percent Treg cells present in the gated CD4+ population. Representative flow cytometric analyses of CD25- and FoxP3-positive (a) T cells alone, and (b) T cells following coculture with ASCs at a 1:1 ratio under low and ambient O2 culture conditions. (c) Representative quantification of percentage of Tregs following coculture of ASCs with T cells. Three independent sets of experiments were performed for each treatment. Data are reported as mean (μ)±SE. ***P<0.001.