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. 2019 Oct 25;9:15296. doi: 10.1038/s41598-019-51773-x

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© The Author(s) 2019

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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Regulation of the Ran protein gradient by TMX2. (A) Flag-TMX2 WT, Flag-isoform 2, or mCherry was overexpressed in HEK293 cells, and the distribution of endogenous Ran was analyzed by immunofluorescence microcopy. (B) The ratio of Ran levels in the nucleus/cytosol of each kind of cell (n = 60) was quantified. (C) si-RNA targeting for the negative control or TMX2 was lipofected into HEK293 cells, and TMX2, importin-β, or Ran protein levels were analyzed by western blotting. (D) Endogenous Ran distribution in TMX2 knockdown cells was analyzed by immunofluorescence microcopy. (E) The ratio of Ran levels in the nucleus/cytosol of each kind of cell (n = 60) was quantified. (F) The nuclear and cytosolic fractions of TMX2-overexpressing or -knockdown cells were extracted, and analyzed by western blotting with anti-Ran antibody. Values are the means ± S.D. for three experiments. The Ran nucleus/cytosol ratio of control cells (mock or si-control) was set at 1.0. *p < 0.05, **p < 0.01.