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. 2018 Apr 30;8:6748. doi: 10.1038/s41598-018-25137-w

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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EGFP-TARG1 rapidly shuttles between the nucleoplasmic and the nucleolar compartment. (a) Fluorescence loss in photo-bleaching (FLIP) experiments in U2OS cells transiently expressing EGFP-TARG1, EGFP-TARG1-G123E or EGFP. mCherry-H2B was co-expressed to define the nucleoplasm (not shown). Representative images are shown. (b–d) EGFP fluorescence intensities of EGFP-TARG1 (upper panels), EGFP-TARG1-G123E (middle panels) and EGFP (lower panels) were measured in each ROI during photo-bleaching of nucleoplasmic EGFP (b), after total photo-bleaching of nucleoplasmic EGFP (c) or during photo-bleaching of cytoplasmic EGFP (d). Fluorescence intensities were normalized to the mean EGFP fluorescence intensity in the same ROI before bleaching and are expressed as mean ± SD of 7–12 cells as indicated.