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. 2017 Jun 15;28(12):1667–1675. doi: 10.1091/mbc.E16-11-0777

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© 2017 Pan et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — Knockdown of sortilin decreases colocalization of myc₇-Glut4 with Vps35 and increases its presence in lysosomes. (A) Total lysates (40 µg/lane) prepared from 3T3-L1 adipocytes stably transfected with myc₇-Glut4 and shRNA against sortilin were analyzed by Western blotting. (B) Top: 3T3-L1 adipocytes stably transfected with myc₇-Glut4 and shRNA against sortilin were stained with goat polyclonal antibody against Vps35 and mouse monoclonal anti-myc antibody, followed by Texas red–conjugated donkey anti-goat IgG and FITC-conjugated donkey anti-mouse IgG. Bottom: 3T3-L1 adipocytes stably transfected with myc₇-Glut4 and shRNA against sortilin were incubated with LysoTracker and stained with mouse monoclonal anti-myc antibody, followed by FITC-conjugated donkey anti-mouse IgG.