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. 2020 Feb 6;10:1984. doi: 10.1038/s41598-019-56877-y

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© The Author(s) 2020

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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Sox2 overexpression induces FN fibrillogenesis and focal adhesion in Schwann cells. (a–c) Graphs show real-time RT-qPCR measurements of Fn1, Fn1EIIIA and Fn1EIIIB mRNA levels in SC^wt and selected clones after 3 days in culture. Gapdh, Ankrd27 and Rictor were used as reference genes, values are relative expression to SC^wt. t-test comparing each clone with the control (N = 5). (d) Representative confocal images of Actin fibres (red) and FN1 (green) immunostainings showing FN expression and fibrillogenesis in SC^wt (arrow head) an in SC^Sox2/Cl2 (arrows) cultured in FN-free medium. Nuclei were counterstained with DAPI (blue). Scale bars, 50 μm and 25 μm. (e) Western blot analysis and quantification of total FN1 protein in the whole lysate of SC^wt and SC^Sox2/Cl2. GAPDH was used as a loading control. (N = 8). (f) pPaxillin immunostaining showing the formation of focal adhesion during the first 60 minutes after seeding on laminin-coated slides. (g) Distribution of adhesion structures during cell spreading, according to their circularity (n ≥ 25 cells/time point). Graphs show mean value ± s.e.m, *p < 0.05, **p < 0.005, ***p < 0.0005.