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. 2017 Jul 14;7:5454. doi: 10.1038/s41598-017-05707-0

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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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Netrin-1 promoted the expression of the oncogene c-Myc. (a) Immunoblotting analyses of the expression of selected oncogenes in U251 and U87MG cells after stimulation with recombinant human netrin-1. (b) Quantification of c-Myc, cyclin D1, p27, cdk2 and cyclin E expression using Gel-Pro Analyzer 4 software. (c) c-Myc expression was detected by real-time PCR in U251 and U87MG cells in the absence or presence of netrin-1. (d) Western blotting analyses of c-Myc in U251 cells that were infected with lentivirus expressing shCtrl or shNetrin-1. GAPDH was used as the loading control. (e) c-Myc expression was detected by real-time PCR in U251 cells that were infected by lentivirus expressing shCtrl or shNetrin-1. (f) Representative images of c-Myc staining in U251-derived tumor sections. Boxed areas in the top row defined regions shown at a higher magnification in the bottom row. Scale bar: 50 µm. (g) Relative expression of c-Myc in U251-derived tumors. The mean IOD of c-Myc was calculated using Image-Pro Plus.