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. 2019 Aug 15;10:258. doi: 10.1186/s13287-019-1367-x

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 1 — iPS cell differentiation. a Graphical summary of experimental groups and design. b Step-wise flow cytometry characterization of hepatocyte differentiation in vitro. Representative image of gated analyses for FOXA2⁺/CXCR4⁺ at day 3, AFP⁺/HNF4A⁺ at day 9, UGT1A1⁺/ALB⁺ at day 18, and phase-contrast image of cellular morphology at day 18. c IF staining for FOXA2/CXCR4 at day 3, AFP/HNF4A at day 9, and UGT1A1/ALB and PAS staining of hepatocytes at day 18 and confluent fibroblast culture PAS staining negative control (n = 3, biological replicates; data displayed as mean ± SEM, bar = 50 μm). d Step-wise flow cytometry characterization of endothelial differentiation in vitro. Representative images of gated analyses for BRACHYURY⁺/GATA4⁺ at day 2, CD34⁺/CD31⁺ at day 6, VECAD⁺/CD31⁺ at day 10, and phase-contrast image of cellular morphology at day 10. e IF staining for CD34 at day 2, VECAD/CD31 and Ac-LDL uptake at day 10, and angiogenesis assay at day 12 (n = 3, biological replicates; data displayed as mean ± SEM, bar = 50 μm). f Step-wise flow cytometry characterization of iNCC-MSC differentiation in vitro. Representative image of gated analyses for P75+/HNK1+ at day 18 and CD73+/CD90+ and IF staining for CD105/F-Actin and phase-contrast image at day 28. g IF staining for P75/VIMENTIN at day 18, CD73/VIMENTIN and CD105 at day 28, and representative images for MSC differentiation assay after 28 days of induction and stained for Alizarin Red, Alcian Blue, and Oil red (n = 3, biological replicates; data displayed as mean ± SEM, bar = 50 μm)