p40^phox accumulates on to the phagosome independent of PI3P. Time-lapse confocal microscopy was used to monitor translocation of coexpressed YFP-p40^phox and Cherry-PX₄₀ in PLB-985 cells during phagocytosis of IgG-zymosan in the absence (A) and presence (B) of 100 nM wortmannin (Videos S6,S7). YFP-p40^phox is detected on the cup, whereas Cherry-PX₄₀ appears after closure (A). Cherry-PX₄₀ does not accumulate in wortmannin-treated cells, although YFP-p40^phox is present on internalized phagosomes (see also Figure 4D). N shows the location of nucleus in the YFP-p40^phox panel. Phagosome stages are indicated and asterisks show the IgG-zymosan phagosomes monitored. Bar represents 5 μm.