Immunofluorescence microscopy of LC3 and LysoTracker in wild-type and <i>CatE</i>^{<i>-/</i><i>-</i>}macrophages.

<p>(<b>A</b>) Cells on glass cover-slips were preincubated with LysoTracker-Red for 30 min, subsequently fixed, permeabilized with 0.3% Tween-20 in PBS, and allowed to react with anti-LC3 antibody. The cells were then incubated with a fluorescence-labeled secondary antibody and visualized by confocal laser microscopy. (<b>B</b>) Based on the data from immunofluorescence microscopy, the number of LC3- or LysoTracker-positive vesicles per cell was counted. The data are shown as percent merged vesicles per cell and acquired from 3 independent experiments. *<i>P</i> < 0.05 for the indicated comparisons.