Inhibition of DNA methylation leads to stable CCR6 expression. (A) Naive CD4⁺ T cells were stimulated with or without proinflammatory cytokines and TGF-β. After 48 hours, Aza was added to the media for another 48 hours. Expression of CCR6 (n > 18, left panel) and methylation of the DMR (n = 3, right panel) were analyzed on day 6. *P < .05. ***P < .001. (B) Naive CD4⁺ T cells were stimulated with Aza and without cytokines as in panel A. After 6 days, cells were sorted according to their CCR6 expression and restimulated under neutral conditions in the absence of Aza for additional 5 days followed by reanalysis of CCR6 expression. Data shown are representative for 2 independently performed experiments.