Figure 5

Regulatory T cells have similar suppressive function, but effector T cells have a lower activation threshold in 5-Aza-treated mice (A) Effector T cells isolated from naive B6 mice were cocultured with Treg cells isolated from 5-Aza-treated (5-Aza Treg) or DMSO-treated (DMSO Treg) EAE mice at disease onset and peak. Treg inhibition assays were then performed by CFSE-labeling. (B) Effector T cells isolated from EAE mice treated with DMSO (DMSO, left panel) or 5-Aza (5-Aza, right panel) were cocultured with Treg cells isolated from 5-Aza-treated mice for Treg inhibition assay (upper panel). For control experiments, effector T cells were stimulated with anti-CD3 antibody only (lower panel). (C) CFSE-labeled CD4⁺CD25⁻ effector T cells isolated from DMSO-treated (Effector T-DMSO) or 5-Aza-treated (Effector T-Aza) EAE mice were cocultured with regulatory T cells isolated from 5-Aza-treated (Treg-Aza) or DMSO-treated (Treg-DMSO) EAE mice for 5 d, and CFSE signals were detected by flow cytometry. A representative CFSE profile is shown. Three independent experiments were performed with similar trends. (D) CD4⁺CD25⁻ T cells isolated from DMSO- or 5-Aza-treated EAE mice were stained with CD69 activation marker before (upper panel) or after (lower panel) stimulation with anti-CD3. Data are representative of three independent experiments with similar results.