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. 2018 Oct 17;50(10):137. doi: 10.1038/s12276-018-0166-2

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© The Author(s) 2018

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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Fig. 2 — a DEGs between TS13-20 spheres (D0) and their differentiated counterparts (D7) were analyzed by RNA sequencing. Genes with expression changes more than four-fold are shown. DEGs are listed in Supplementary Table S2. b The cholesterol biosynthesis-related pathway was the dominant pathway altered in sphere (D0) cells according to functional enrichment analysis using FunRich (http://www.funrich.org). The top five pathways are presented. c Down- and upregulated DEGs were subjected to gene network analysis using Cytoscape (http://www.cytoscape.org). Gene networks with P values < 0.005 are shown. d The change in the mRNA expression of various metabolic pathway genes between D0 and D7 cells was analyzed. Cholesterol-related genes were the only metabolic genes that varied between the two samples. The marker size is proportional to the log2-fold change (D0/D7). e Heatmap showing a clear distinction between the cholesterol pathway-related gene expression levels in the two samples. f Changes in the expression of cholesterol pathway genes were validated by RT-PCR and real-time quantitative PCR. The data are presented as the means of three independent experiments. ***P < 0.001 relative to D0