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. 2016 Mar 14;212(6):677–692. doi: 10.1083/jcb.201507112

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© 2016 Johnson et al.

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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 8. — Assessment of lysosomal mobility by photoactivation. (a) Lysosomes were loaded with Alexa Fluor 647–dextran (purple) and CMNB-caged carboxyfluorescein-dextran. A ROI (indicated by solid white circle) was selectively illuminated using a 405-nm laser to photoactivate the CMNB-caged carboxyfluorescein-dextran (green in middle and right panels). The resulting fluorescence was imaged using a 488-nm laser every 1 min for 30 min or every 4 min for 2 h. Photoactivated lysosomes were tracked using Imaris software (right). (b) Lysosomes were tracked in cells where CMNB-caged carboxyfluorescein-dextran was photoactivated in either the cell center or the cell periphery. The total track length over 30 min was quantified. Error bars represent SEM (n = 104–204 lysosomes from three to five cells from five independent experiments). (c) The net displacement over 2 h was quantified. Error bars represent SEM (n = 137–285 lysosomes from three to seven cells from two independent experiments).