Translocation of YFP-tagged p67phox and p40phox during phagocytosis of IgG-zymosan in the presence and absence of wortmannin. Time-lapse confocal microscopy was used to monitor phagocytosis of IgG-zymosan by PLB-985 neutrophils expressing YFP-tagged p67phox or p40phox (Videos S1Video 2. Time-lapse movie of PLB-YFPp40 cells corresponding to the sequence shown in Figure 4B (MOV, 598 KB)Video 3. Time-lapse movie of wortmannin-treated PLB-p67YFP cells corresponding to the sequence shown in Figure 4C (MOV, 239 KB)–S4). N shows the location of nucleus and asterisks indicate the IgG-zymosan phagosomes monitored. Bar represents 5 μm. (A,B) PLB-p67YFP and PLB-YFPp40 cells. The relative fluorescence intensity compared with the cytosol for 5 phagosomes at indicated stages is shown in the graphs, as mean plus or minus SE. Internalized indicates 200 or more seconds after phagosome closure. (C,D) PLB-p67YFP and PLB-YFPp40 neutrophils pretreated with 100 nM wortmannin before incubation with IgG-zymosan. N shows the location of nucleus and asterisks indicate the IgG-zymosan phagosomes monitored. Bar represents 5 μm. The relative fluorescence intensity compared with the cytosol for 5 to 7 phagosomes at indicated stages is shown in the graphs, as mean plus or minus SE. Internalized indicates 120 or more seconds after phagosome closure.