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. 2020 Oct 7;10:16714. doi: 10.1038/s41598-020-73698-6

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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ACR interferes with genes that are important for neuronal differentiation during retinoic acid- induced SH-SY5Y cell differentiation. (a) Heatmap of the gene expression changes of several neuronal markers and other genes of interest (Day 3, Day 6 and Day 9 versus undifferentiated cells—RNA-sequencing input data) and following exposure to ACR (1 and 70 µM, after 9 days of differentiation versus untreated cells at the same time point—PCR input data). Colour coding refers to the log2(fold change). Genes are ranked according to decreasing log2(fold change) of the contrast Day 9 versus undifferentiated cells. (b) BDNF protein levels after 9 days of differentiation and exposure to 1 and 70 μM of ACR by western blot (image for one representative gel is included). The data from the protein levels were normalized against α-tubulin and presented as the mean ± SEM (N = 4). Results were analysed using one-way ANOVA followed by Dunnett’s multiple comparisons test. *p ≤ 0.05 compared to control (cells exposed to differentiation medium without ACR). Full length blots are presented in Supplementary Figure S4 online.