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Evaluation of LC3 and ATG13 dynamics in AS-stimulated microglial cells using live-imaging.
Evaluation of LC3 and ATG13 dynamics in AS-stimulated microglial cells using live-imaging. (A,B) BV2 cells stably expressing GFP–LC3 (green) were stimulated with fAS (red) for 12 h. Imaging was performed at 1 frame per 10 s for 1 h and a selected interval within this sequence for two experiments is shown. LysoTracker Blue was added 30 min previous to image acquisition. Note that autophagosomes form a ring-like structure around LysoTracker+/fAS+ structures. (C,D) BV2 cells stably expressing GFP–ATG13 (green) were transfected with CFP–LC3 (red) and stimulated with fAS (blue) and imaged as described above. Arrowhead indicates the first discernible ATG13 punctum during autophagosome formation (C) and ATG13 forming a puncta pattern similar to LC3 (D). Scale bars: 2 µm. Claudio Bussi et al. J Cell Sci 2018;131:jcs226241 © Published by The Company of Biologists Ltd
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