Immunofluorescence microscopy of LC3 and LysoTracker in wild-type and CatE-/-macrophages.
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posted on 2013-12-05, 04:51 authored by Takayuki Tsukuba, Michiyo Yanagawa, Tomoko Kadowaki, Ryosuke Takii, Yoshiko Okamoto, Eiko Sakai, Kuniaki Okamoto, Kenji Yamamoto(A) 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) 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. *P < 0.05 for the indicated comparisons.
