FIG. 6.
Effect of K252a, PP2, and AG1478 on phosphorylation of PDK1 induced by alpha-toxin in rabbit neutrophils. (A) Rabbit neutrophils (1.0 × 106 cells/ml) were preincubated with various concentrations of K252a, PP2, or AG1478 at 37°C for 60 min, and the cells were incubated with 1.0 μg/ml alpha-toxin at 37°C for 30 s. The extracted phospho-PDK1 was subjected to SDS-PAGE and Western blotting using antibodies for PDK1 and phospho-PDK1. A typical result from one of five experiments is shown. (B) Rabbit neutrophils (1.0 × 106 cells/ml) were preincubated with various concentrations of K252a, and the treated cells were incubated with or without alpha-toxin at 37°C for 10 min. Intracellular DG levels were determined as described in Materials and Methods. Values represent means ± standard error for five to six experiments.