FIGURE 1.

DUSP16 is differentially expressed in Th clones. A, 28-4, a Th1 clone, and MS-SB, a Th2 clone, were starved overnight without growth factors and either left untreated or treated with 1 μg/ml LPS for indicated times. Total RNA was extracted for Northern blot analysis using a ³²P-labeled cDNA DUSP16 probe. The blot was stripped and rehybridized with a mouse PAC-1, MKP-1, M3/6, T-bet, or β-actin cDNA probe. B, 28-4 and MS-SM Th clones were starved and left untreated or treated as described in A. Total cellular lysates were prepared, and protein levels of DUSP16 were analyzed by Western blotting using anti-DUSP16 polyclonal antibody. As a control, 10% of each lysate was used to detect α-tubulin using anti-α-tubulin mAb. C, 28-4 and MS-SB Th clones were starved as described in A, and cells were left untreated (unstim) or treated for 4 h with IL-2 (10 ng/ml), IL-12 (10 ng/ml), IL-4 (10 ng/ml), or LPS (1 μg/ml). Total RNA was extracted, and gene expression of DUSP16 was analyzed. The ethidium bromide-stained gel is shown.