Immunoblot analysis of the effect of EBV on cPLA₂ phosphorylation in enriched monocyte suspensions. Enriched monocyte suspensions (5 × 10⁶ cells/0.1 mL) were preincubated or not in the presence of EBV for 30 minutes at 37°C, diluted 10-fold with HBSS, and subsequently stimulated with fMLP (1 μmol/L for 10 minutes) or A23187 (50 nmol/L for 5 minutes). Incubations were stopped on ice and whole cells were processed for SDS-PAGE analysis and immunoblotting of cPLA₂ as described in the Materials and Methods. The upper and lower bands of the doublet represent the phosphorylated (cPLA₂-P) and nonphosphorylated (cPLA₂ ) forms of the cPLA₂ , respectively. (A) The data obtained using fMLP and A23187 as second stimuli are derived from separate experiments. (B) Enriched monocyte suspensions (3 × 10⁶ cells/0.1 mL) were preincubated or not in the presence of EBV for the indicated times at 37°C, diluted 10-fold with HBSS, stimulated with A23187 (50 nmol/L for 5 minutes), and processed for analysis of cPLA₂ as described above. Experiments shown are representative of three performed.