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. 2019 Jul 19;9:10506. doi: 10.1038/s41598-019-46848-8

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© The Author(s) 2019

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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Treatment with a high-dose GSK3β inhibitor predominantly induced posterior DE. (A) A schematic diagram of posterior DE induction using Activin A and CHIR99021. (B) An expression analysis of SOX17 and CDX2 in differentiated cells treated with the indicated dose of CHIR by qPCR (n = 3 independent experiments; mean ± SEM). (C) Immunostaining of SOX17 and CDX2 in differentiated cells. Representative fluorescence microscopic images for treatment with 1 µM (upper panels) and 5 µM CHIR (lower panels) are shown. Scale bar, 50 μm. (D) The proportion of CDX2-positive cells after treatment with 1 or 5 µM CHIR (n = 3 independent experiments; mean ± SD). *p < 0.05; two-tailed paired t-test. Abbreviations: iPSc, induced pluripotent stem cell; DE, definitive endoderm; CHIR, CHIR99021.