Fig. 5. Cholesterol depletion disrupts PtdIns(3,4,5)P₃ accumulation. (a) Distribution of filipin staining in fixed neutrophils before and after treatment with methyl-ß-cyclodextrin. (b) Quantification of the reduction in `peripheral' or membrane staining in neutrophils before and after treatment with methyl-ß-cyclodextrin. The data are shown as the percentage of filipin staining at the cell periphery. (c) Images of a neutrophilic HL60 cell treated with methyl-ß-cyclodextrin demonstrating the failure of GFP-Akt-PH to accumulate at phagocytic cups. The cell was stained with DiIC₁₆(3) (red) and GFP-Akt-PH (green) with the position of two zymosan particles indicated. This cell was typical of at least ten others. Bars, 4 µm.