Fig. 5.

The mutant EGFP-4KE/A-cPLA₂α does not translocate to the phagosome in human macrophages. A: Human macrophages transfected with the mutant EGFP-4KE/A-cPLA₂α were subjected to synchronized phagocytosis with Alexa Fluor 594-labeled opsonized zymosan for the indicted times, fixed and analyzed by confocal microscopy. Framed phagosomes have been enlarged and the intensity of the fluorescence analyzed by pseudocolor (left upper inserts at 15 and 30 min). B: Cells transfected with the construct EGFP-cPLA₂α or EGFP-4KE/A-cPLA₂α were subjected to a frustrated phagocytosis assay and plated over noncoated glasses (Ctrl) or IgG- coated glasses (IgG) as indicated, and analyzed by confocal microscopy. Pictures of the XZ axis of the cells were taken and the intensity of the fluorescence obtained was analyzed in pseudocolor. The figure is representative of more than 40 cells that were analyzed per experiment and the experiment was repeated four times. White bar = 10 μm.