Figure 5

Up-regulation of Foxp3 expression in iTreg by GCSE treatment. CD4+ T cells isolated from the spleen and lymph node of 8 week old Foxp3-GFP knock in mice were stimulated in a medium supplemented with anti-CD3 (1 μg/ml) / CD28 Ab (3 μg/ml), anti-IL-4 Ab (10 μg/ml), anti-INF-γ Ab (10 μg/ml) and TGF-β (5 ng/ml) at day 1 and additional 50 U/ml rhIL-2 at day 3. Then iTreg cells were stimulated with various concentrations of GCSE in the presence of PMA (50 ng/ml)/ ionomycin (1 μM) for 12 hrs. (A) Relative expression levels of Foxp3 of GCSE treated samples were compared with control sample by qRT-PCR. Expression level of HPRT was used as an internal control. (B) GFP signal was measured as protein level of Foxp3 by flow cytometry. Error bars indicate SD. One (*), two (**), and three (***) indicate p < 0.05, p < 0.01, and p < 0.001 respectively. Data are representative of three independent experiments.