FIG 3.

Role of MKK3/6 in GRA-mediated p38 activation. (A and B) Infected cells as stated in the legend to Fig. 1 were either treated with GRA (20 μM) (A) or left untreated (B) for different times, and the total and phosphorylated levels of various MAP kinases were detected by Western blotting using the respective anti-MAP kinase antibodies. (B, C, and D) Cells were transiently transfected with the wild-type or dominant-negative constructs of MKK3 (B), the wild-type or dominant-negative constructs of MKK6 (C), and the wild-type or dominant-negative constructs of both MKK3 and MKK6 (D) for 24 h. Thereafter, they were infected and treated with GRA for 4 h, and in all the samples, the levels of total and phosphorylated p38 were determined by Western blot analysis. The bands were analyzed densitometrically, and the fold changes are indicated below each blot. All experiments were repeated at least three times each, and one set of representative data is shown.