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. 2019 Jan 10;9:30. doi: 10.1038/s41598-018-37188-0

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© The Author(s) 2019

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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Long-term treatment with PLX4032 and dacarbazine induces cross-resistance via MAPK and PI3K/AKT/mTOR hyperactivation in more aggressive and invasive T24.6.9 melanoma cells. Parental cells (T24.6.9) in comparison to twelve months chronically treated cells (T24.6.9-10Da, T24.6.9-1cP and T24.6.9-0.1PLX) were analysed. (A) mRNA-expression levels of genes related to marker expression and cell migration (left) and chemoresistance (right). Values are normalised to housekeeping gene GAPDH and parental cells [=1.0]. Analysis of pathway signalling by protein (B) immunoblots and (C) quantification. Values are normalised to housekeeping gene GAPDH and parental cells [=100%]. The blots were cropped to focus upon the specific proteins indicated. The entire gels blots are shown in Supplementary Figs S6 and S7. (D) Relative activation (left) and expression (right) of ERK 1/2 during PLX4032 treatment of resistant 0.1PLX cells. Values are normalised to housekeeping gene GAPDH and the untreated, PLX-resistant control [=100%]. (n values depicted in figure; mean ± s.e.m.; *P < 0.05; **P < 0.01; ***P < 0.001) 10Da = long-term 10 µM dacarbazine-treated cells; 1cP = long-term 1 µM cisplatin-treated cells; 0.1PLX = long-term 0.1 µM PLX4032-treated cells; p = phosphorylated.