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. 2019 Apr 5;9:5740. doi: 10.1038/s41598-018-36456-3

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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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Regulation of ETV5 expression through MAPK/ERK. (a) ETV5 mRNA and (b) ETV5 and pERK1/2 protein levels in cells overexpressing wildtype FGFR3 stimulated for 5 hours with FGF1, with or without pre-treatment with the MAPK inhibitor U0126. ‘Control’ are unstimulated cells, ‘+ FGF1’ are cells stimulated with FGF1, ‘U0126’ are unstimulated cells treated with U0126, and ‘U0126 + FGF1’ are cells stimulated with FGF1 following 1 hour pre-treatment with U0126; (c) Expression of MAPK/ERK regulatory genes in TERT-NHUC with overexpressed mutant (S249C) FGFR3, compared with control cells transduced with the empty vector (Control) and cells with overexpressed S249C FGFR3 but silenced ETV5 (S249C shETV5). mRNA expression levels were relatively quantified using Taqman Real-Time RT-PCR with SDHA as internal control, while ETV5 and pERK1/2 proteins were visualized by western blotting with specific antibodies using alpha-tubulin as loading control. All experiments were repeated in triplicate. ETV5 silencing was performed using the sh155 hairpin. ‘*’ indicates a statistical significant difference between samples.