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. 2001 Feb 19;152(4):657–668. doi: 10.1083/jcb.152.4.657

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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LC3 does not target to membrane in APG5⁻/− cells. Wild-type ES cells (A–C) and APG5⁻/− cells (D and E) were cultured in Hanks' solution for 2 h and fixed. Localization of LC3 was examined by silver-enhanced immunogold electron microscopy using an antibody against recombinant LC3. The isolation membrane (arrow), autophagosomes (open arrowheads) and autolysosome (closed arrowhead) are indicated. Bar, 1 μm. (F) Total lysates from APG5 ^+/+ and APG5 ^−/− cells and their various stable transformants cultured in Hanks' solution for 2 h were subjected to immunoblot analysis with anti–LC3 antibody. Positions of LC3-I and LC3-II are indicated.