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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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FGF10 enhanced urothelial differentiation and stratification. (A) After posterior DE induction, cells were treated with 0, 100, 200, or 500 ng/ml FGF10 from days 3 to 18. (B) A quantitative RT-PCR analysis of urothelial markers (UPK Ia, UPK II, and UPK III) in differentiated cells at day 18 after treatment with the indicated concentrations of FGF10. Data are shown as the mean ± SEM (n = 6 independent experiments). *p < 0.05; Dunnett’s test. (C) Immunostaining of UPK III, CK20, p63, ZO-1 and E-Cadherin in differentiated cells. Scale bar, 50 µm. (D) Phase contrast images of differentiated cells with (right panel) or without (left panel) FGF10 treatment (100 ng/ml). (E) Optical coherence tomography (OCT) images of differentiated cells treated with (lower panel) or without (upper panel) FGF10 in high-resolution mode. (F) The thicknesses of the differentiated cells with (right panel) or without (left panel) FGF10 treatment were visualized using a color-scale heatmap from 10 to 70 μm in low-resolution mode. Scale bar, 1 mm. (G) A histogram of the OCT analysis of differentiated cells with (red line) or without (blue line) FGF10 treatment in low-resolution mode. (H) An EdU assay for the cells with and without FGF10 treatment. Representative fluorescence microscopic images of EdU (left panel) and the proportions of EdU-positive cells (right panel) are shown (n = 5 different fields; mean ± SD). *p < 0.05; two-tailed paired t-test. Abbreviations: iPSc, induced pluripotent stem cell; DE, definitive endoderm; CHIR, CHIR99021; BPE, bovine pituitary extract; ATRA, all-trans retinoic acid; TZ, troglitazone; PD, PD153035; UPK, uroplakin.