FIGURE 3.

Effects of reagents to regulate RhoA and Rap1 activities on the binding of SOZ particle. Raw264.7 cells (2 × 10⁵) were pretreated with 50 μm 8CPT-2Me-cAMP, 10 μg/ml Tat-C3 toxin, 10 μm LPA, 5 μm RhoA activator I (CN01) for 60 min at 37 °C. FITC-conjugated zymosan particle (2 × 10⁶) were applied for 30 min on ice for particles to bind to cells, and then unbound F-SOZ particles were washed out three times with cold PBS. Total fluorescence of SOZ particles bound to cells after washing was measured by a fluorescence spectrophotometer. At this step, crystal violet could completely quench the fluorescence of FITC, demonstrating that phagocytosis is almost blocked, and all F-SOZ particles were bound to the surface of cells. Data are presented as mean ± S.E. (error bars) of five (A) or three (B) independent experiments.