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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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Binding between TMX2 and importin-β or Ran. (A) Scheme of GST-tagged TMX2 full-length and deletion mutants. (B,C) GST-tagged TMX2 proteins were immobilized on glutathione-Sepharose beads and incubated with purified His-tagged importin-β or Ran protein. The precipitated importin-β or Ran was analyzed by immunoblot with anti-His tag antibody. (D) TMX2-packed glutathione-Sepharose was incubated with HEK293 cell lysates, and the precipitant with TMX2 was analyzed with anti-importin-β or Ran antibody. (E) Flag-importin-α, -importin-β, -CRM1, or -RanQ69L was expressed in HEK293 cells, and cell lysates were immunoprecipitated with anti-TMX2 antibody. The precipitates were detected with anti-Flag antibody. (F) The ratio of band intensity of binding/input was quantitated. Values are the means ± S.D. for three separate experiments. The value of importin-β was set at 1.0.