FIGURE 5.

Altered CD4⁺ T cell differentiation in T cell-specific dusp16 Tg mice. A, flow cytometric analyses of peripheral T cells for control C57BL/6, dusp16^WT Tg, and dusp16^C244S Tg mice. Splenic cells were prepared from 6-week-old mice and stained with phycoerythrin-conjugated anti-CD8α and FITC-conjugated anti-CD4 mAb. The percentages of quadrant fractions are indicated. These data represent one of several repeated experiments. B, purified CD4⁺ T cells from control C57BL/6 or T cell-specific dusp16 Tg mice differentiated under the Th1 or Th2 condition. Cell lysate was prepared on day 3 for Western blotting with the indicated antibodies. C, purified CD4⁺ T cells from control C57BL/6 or T cell-specific dusp16 Tg mice differentiated as in B. Total RNA was extracted on the indicated days, and gene expression of IFNγ, IL-4, GATA-3, and β-actin was analyzed by semiquantitative RT-PCR. D, day 7. The differentiated cells in B were collected and restimulated with 2 μg/ml plate-bound anti-CD3 mAb for 24 h. Supernatants were analyzed for IFNγ or IL-4 by ELISA. Data are expressed as mean ± S.D. (error bars) of three independent experiments.