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. 2017 Jul 5;7:4695. doi: 10.1038/s41598-017-04835-x

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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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Effect of pBMP-9 and SpBMP-9 on the differentiation of SH-SY5Y cells. (A) Western blots of MAP-2 and βactin (2 independent experiments) and densitometric analyses of MAP-2 bands normalized to βactin (means ± SEM) for SH-SY5Y cells stimulated with 0.1 nM BMP-9, pBMP-9 and SpBMP-9 +/− 10 μM RA for 3d and 5d (*p < 0.05, **p < 0.01). Only cropped pictures of western blots showing the 80 kDa MAP-2 isoform were presented in order to allow a better comparison between experimental conditions. Complete gel pictures are available in the supplementary data file. (B) Merged pictures showing immunostaining for neuronal differentiation markers βIII-tubulin (Alexa Fluor® 488, green), MAP-2 (Alexa Fluor® 488, green), NeuN (FITC, green) and NSE (FITC, green), and nuclei staining (Hoechst, blue) of SH-SY5Y cells stimulated for 5d with 0, 0.1 or 1 nM BMP-9, pBMP-9 and SpBMP-9 +/− 10 μM RA. Pictures are representative of at least two independent experiments (Bar = 100 μm).