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. 2015 Dec 1;26(24):4427–4437. doi: 10.1091/mbc.E15-04-0207

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© 2015 Tewari 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 1: — Dominant-negative GGA1 blocks Mn-induced GPP130 trafficking. (A) Cells were transfected with GFP-tagged GGA1 or GGA1-DN and then either left untreated (Ø) or treated with Mn for 5 h. Images show GFP fluorescence from the transfected proteins and anti-GPP130 staining, with the final panel using uniform thresholding of the GPP130 image to aid visualization of small punctae. Note that GGA1-DN blocked GPP130 appearance in punctae. Bar, 5 μm. (B) Cells expressing GGA1 or GGA1-DN were treated with monensin for 30 min to induce GPP130 trafficking to endosomes. (C) Quantified appearance of GPP130 in peripheral punctae of cells expressing GGA1 or GGA1-DN after Mn or monensin (Mon) treatment (mean ± SEM, n = 3, >10 cells/experiment, *p < 0.05). (D) Total cell fluorescence of GPP130 in cells expressing GGA1 or GGA1-DN before or after Mn (mean ± SEM, n = 3, >10 cells/experiment).