FIG. 2.

Activation of the 63- and 69-kDa Paks in neutrophils treated with different concentrations of agonists. Neutrophils were stimulated with different amounts of the indicated agonists for 15 s, and the 63- and 69-kDa Paks were assayed as described in Materials and Methods. In lanes a and b of all panels, cells were also treated for 15 s with 0.25% (vol/vol) Me₂SO (i.e., unstimulated cells) and 1.0 μM fMLP for comparative purposes, respectively. (A) The concentrations of PAF in lane a and in lanes c through h were 0.0 μM (0.25% [vol/vol] Me₂SO), 5.0 μM, 1.0 μM, 0.10 μM, 50 nM, 10 nM, and 1.0 nM, respectively. (B) The concentrations of LTB₄ in lanes c through i were 0.0 μM (0.25% [vol/vol] ethanol), 0.50 μM, 0.10 μM, 20 nM, 5.0 nM, 1.0 nM, and 0.10 nM. (C) The concentrations of IL-8 in lanes c through i were 0.0 μM (0.001% [wt/vol] BSA in PBS), 0.25 μM, 0.125 μM, 12.5 nM, 6.3 nM, 1.3 nM, and 0.13 nM. (D) The concentrations of RANTES in lanes c through i were 0.0 μM (0.001% [wt/vol] BSA in PBS), 0.13 μM, 65 nM, 13 nM, 6.5 nM, 1.3 nM, and 0.65 nM. (E) The concentrations of sulfatide in lanes c through i were 0.0 μM (PBS), 400 μg/ml, 200 μg/ml, 100 μg/ml, 50 μg/ml, 10 μg/ml, and 1.0 μg/ml. Positions of the 63- and 69-kDa Paks are designated by arrows and arrowheads, respectively.