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. 2017 Aug 22;7:9166. doi: 10.1038/s41598-017-09558-7

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© The Author(s) 2017

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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Akt activation might be involved in the Nm23-H1-mediated IRES-translational activity of Sp1 mRNA. (A) Samples were harvested from H1299 and CL1-5 cell lines treated with Akt inhibitors, Ly294002 and MK2206, and were analyzed by Western blotting with antibodies against the indicated proteins (a). After three independent experiments, the proteins levels were quantified and statistical analysis was done with a t-test, *p < 0.05; **p < 0.01; ***p < 0.005 (b). (B–F) Samples harvested from H1299 cells with overexpression of GFP, GFP-Nm23-H1, GFP-Nm23-H1(S120A) or GFP-Nm23-H1(S122A) were analyzed by Western blotting with antibodies against the indicated proteins and real time PCR (B), 5′UTR-driven luciferase activity (C), RNA-IP with anti-GFP antibodies (D), cell counting (E) and a colony-formation assay (F). After three independent experiments, the proteins levels were quantified and statistical analysis was done with a t-test, *p < 0.05; **p < 0.01; ***p < 0.005. (G) The proposed model shows that Nm23-H1 positively regulates Sp1 expression through an increase in its translational activity in a cap-independent manner.