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. 2011 Oct 24;6(10):e26626. doi: 10.1371/journal.pone.0026626

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Humphries et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Confocal microscopy image of ECFP-Rab7 (blue), LAMP1-EYFP (green), and an antibody against M6PR labeled with a Cy5-labeled secondary antibody (red). Smaller images show a M6PR-positive (red) Rab7-vesicle (blue, top) and a M6PR-negative LAMP1-vesicle (green, bottom). (B) A large fraction of Rab7- and Rab7/LAMP1-vesicles are positive for M6PR; 57±7% and 63±15%, respectively. A smaller fraction of LAMP1-vesicles are positive for M6PR, 23±10%. Error bars show standard deviations. P-values<0.001 are indicated by ***. N.S. indicates a p-value >0.05. The graph shows the analysis of 10–20 of each type of vesicle per cell in 10 cells from 4 distinct experiments. Similar results were obtained for BS-C-1 cells stably expressing ECFP-Rab7 and for HeLa cells (Figure S4). Unmerged images are shown in Figure S5.