Fig 4. Effect of DHA on Tregs proliferation and induction in vitro.

CD4⁺CD25⁺ (Tregs) and CD4⁺CD25^- (Teffs) were isolated from spleen of wild type mice by MACS. A) Effect of DHA on nTregs proliferation. Purified CD4⁺CD25⁺ Tregs (5x10⁴/well) were activated with anti-CD3/CD28 beads and IL-2 (100U/ml) in the presence or absence of varying dose of DHA (6.25–50μM) for 3d. Mean (±SD) results were graphed, *p<0.05,**p<0.01. B) Effect of DHA on Tregs induction. Teffs were incubated (1x10⁵/well) in flat-bottom 96-well plate coated with anti-CD3 Ab (5μg/ml) in the presence of soluble anti-CD28 Ab (2μg/ml) with or without TGF-β1 (2μg/ml) and DHA (25μM). Representative histogram of Foxp3⁺cells (gated from CD4⁺CD25⁺cells) was shown.