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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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Nm23-H1 and hnRNPA2/B1 overexpressed in lung cancer are recruited to the 5′-UTR of Sp1 mRNA. (A) In vitro transcribed pGEM-Sp1-5′-UTR RNA was used as a probe and was incubated with the total cell lysates of HeLa cells. The pulldown mixtures were analyzed using sliver staining and LC MS/MS. (B) Samples were harvested from cells with GFP-Nm23-H1 or myc-hnRNPA2/B1 overexpression for IP with antibodies against GFP or myc. RNA fragments were isolated from IP samples for RT-PCR. (C) Samples were collected from cells with GFP or GFP-Nm23-H1 expression for IP with anti-Nm23-H1 (a) or anti-GFP (b). IP samples were analyzed by Western blotting with antibodies against the indicated proteins. (D) The levels of Sp1, Nm23-H1and hnRNPA2/B1 in the various cancer cell lines were studied by Western blotting with antibodies against indicated proteins. (E,F) The levels of Sp1, Nm23-H1and hnRNPA2/B1 in mice with Kras^G12D- and EGFR^L858R-induced lung cancer were studied using IHC staining (E) and Western blotting (F). After three independent experiments, the levels were quantified and statistically analyzed with t-test, *p < 0.05; **p < 0.01; ***p < 0.005.