FIG.1.

Phosphorylation of PKC isoforms on treatment of rabbit neutrophils with alpha-toxin. (A) Rabbit neutrophils (1.0 × 10⁶ cells/ml) were incubated with 1.0 μg/ml alpha-toxin at 37°C for various periods, and PKC isoforms extracted by 7.5% trichloroacetic acid were subjected to SDS-PAGE and Western blotting using specific antibodies for phosphorylated PKCs. A typical result from one of five experiments is shown. (B) Rabbit neutrophils (1.0 × 10⁶ cells/ml) were preincubated with various concentrations of rottlerin or calphostin C at 37°C for 60 min. The cells were incubated with or without 1.0 μg/ml alpha-toxin at 37°C for 15 min. Production of superoxide was monitored for 15 min based on MCLA chemiluminescence. The integrated data (area under the curve) are plotted. The generation of O₂⁻ induced by alpha-toxin alone was set as the maximal response (100%), against which all other results were compared. *, P < 0.01. Data shown are the mean ± standard error from three different experiments.