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Fig. 7. Inhibition of Phagocytic cup closure by PI 3-kinase inhibition and cholesterol depletion. The failure to complete phagocytosis and the distribution GFP-Akt-PH (green) is shown after treatment with LY294004 (a) and methyl-ß-cyclodextrin (b). The incomplete phagocytic cups are indicated in a by arrows and the position of a single zymosan particle indicated in b by the white circle. A plot of the intensity of peripheral GFP-Akt-PH is shown below b with the position of the incomplete phagocytic cup indicated by the double-headed arrow. (c) Histograms of completed phagocytosis (darker shading) and the formation of phagocytic cups (lighter shading) 20 minutes after micromanipulated delivery of iC3b-opsonised zymosan particles to (i) neutrophils and (ii) neutrophilic HL60 cells untreated (control) or treated with LY294004 (LY) and methyl-ß-cyclodextrin (MßCD) as a percentage of deliveries. There was no significant difference between treatments in the total of incomplete and complete phagocytosis (both shadings in a single histogram column) showing that binding and phagocytic cup formation were not affected by the treatments, whereas the percentage of failed phagocytosis which arrested at the stage of phagocytic cup formation (lighter shading) was increased by the treatments. Bars, 3 µm.
