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Fig. 6. Eosinophil granules are apparent intracellular sites of acidification. Purified eosinophils were loaded with LysoSensor Yellow/Blue DND-160 and stimulated with media control, PAF (3 µM) or IL-5 (0.2 µM) at room temperature (n=10-15 images from four individual isolations for each condition). Confocal microscopy was used to document fluorescence changes under each condition and overlays of the blue and yellow fluorescence of each image are shown. Original images were taken at a magnification of x630. (A) Low power confocal images of eosinophils stimulated with PAF for varying amounts of time (0-15 minutes). (B) Representative images of eosinophils stimulated with IL-5 or PAF for 40 minutes at room temperature. Note the granular appearance of the dye, consistent with accumulation of the dye within the characteristic secondary granules of the eosinophil.





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