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. 2018 Jan 12;8:583. doi: 10.1038/s41598-017-19073-4

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© The Author(s) 2018

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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Activation of the IGF-IR, AKT and ERK1/2 signaling intermediates. Total protein from WM35 (a) and Sk-MEL28 (b) cells exposed to VN, either alone or in combination with increasing concentrations of IGF-I and IGFBP-3, was collected at the indicated time points and assessed by Western blot. Blots were probed for phosphorylated IGF-IR, AKT and ERK1/2 and subsequently stripped and re-probed to determine total levels of IGF-IR, AKT and ERK1/2.