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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 regulates Sp1 and hnRNPA2/B1. (A–C) Nm23-H1 (A,B) or hnRNPA2/B1 (C) was knocked down in CL1-5 cells; thus, the mRNA and protein samples were harvested for real time-PCR and Western blotting with antibodies against indicated proteins. After three independent experiments, the levels of the proteins and mRNA were quantified and statistically analyzed with a t-test, *p < 0.05; **p < 0.01. (D) The pRF and pRSp1F plasmids were transferred into lung cancer cells with Nm23-H1 knockdown (a) or GFP-Nm23-H1 overexpression (b). After 24 h of incubation, cell extracts were collected for a luciferase activity assay and analyzed with t-test, *p < 0.05; ***p < 0.005. (E) Cells with or without knockdown of Nm23-H1 were treated with cyclohexmide. Samples harvested at the indicated time points were analyzed by Western blotting with antibodies against hnRNPA2/B1, Nm23-H1 and tubulin (a). After three independent experiments, the protein level of hnRNPA2/B1 was quantified (b). Samples were harvested from H1299 cells with or without GFP-Nm23-H1 expression treated MG132 (10 μM) for IP with anti-hnRNPA2/B1 antibodies, and then samples were analyzed by Immunoblotting assay with indicated antibodies (c).