FFO attenuates mature Th1/2 cell functions in experimental atopic dermatitis. (A) Mice were stimulated with DNCB for up to 31 days at 2-day intervals, given 100 mg/kg FFO or NFO every day, and were painted with hydrocort every other day, as described in Figure legend 1. After sacrifice, the spleen was isolated and photographed to record the morphologic alteration. (B) Splenocytes were isolated from mice in each experimental group and 1.0 × 106 cells/mL were stimulated with anti-CD3 (1 μg/mL) and anti-CD28 (0.5 μg/mL) for 3 days. Mouse IL-4 and IFN-γ were measured in the culture supernatant by ELISA. (C) Cells were permeabilized with T-bet fixation/permeabilization buffer and stained with anti-T-bet-Alexa Fluor® 488 and anti-GATA3-PE and analyzed by FACS. Data are representative of 8 mice per group. The measurements were made in triplicate and are shown as mean ± S.D (n = 8 mice per group). (B) *P < 0.05; **P < 0.005; and *** P < 0.001 compared to mice sensitized with DNCB alone (induction group).