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. 2008 Jun 11;17(17):2723–2737. doi: 10.1093/hmg/ddn174

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Published by Oxford University Press 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Increased autophagy in fibroblasts from MLIV patients. (A) Control fibroblasts grown in complete media (untreated, left), starved in EBSS media for 3 h (starvation, center) or starved for 3 h followed by 1 h of recovery in complete media (recovery, right), were fixed, permeabilized and immunostained with a polyclonal antibody to endogenous LC3. (B) LC3 staining in untreated control and MLIV fibroblasts. Note that in contrast to a diffuse staining in control fibroblasts, LC3 shows a vesicular staining in MLIV fibroblasts. (C) MLIV fibroblasts grown in complete media were fixed, permeabilized and immunostained with a polyclonal antibody to LC3 (red) and a monoclonal antibody against CD63 or p62 (green). Insets show a 3-fold magnification of the indicated region (D) Double immunostaining of untreated MLIV cells with anti-LC3 (green) and anti-ubiquitin (red). Cells were examined by confocal fluorescence microscopy. Merging the images in the red and green channels generated the third picture on each row; yellow indicates overlapping localization. Scale bars = 10 µm.