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. 2019 Jan 29;9:913. doi: 10.1038/s41598-018-37485-8

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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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Stepwise TF cocktail administration induces nephrogenesis. (a) Schematic of the differentiation protocol from hESCs in 2D culture. (b) Relative mRNA expression levels of PAX2, PAX8, LHX1, WT1, HNF1B and POU5F1 differentiated from hPSCs. (c) Immunocytochemistry for PAX8, LHX1, markers of pretubular aggregate, on day 4 in cells differentiated from hPSCs using protocol depicted in. Scale bar, 200 μm. (d) Flow cytometry for PAX8, LHX1 in hESCs on day 4. Samples stained with secondary antibodies alone were used as controls. (e) Immunocytochemistry for CDH6, LHX1, PAX8 and JAG1, markers of renal vesicle, on day 8 in cells differentiated from hPSCs using protocol depicted in. Scale bar, 100 μm. (f) Immunocytochemistry for proximal tubule (CDH2, AQP1, CDH16), distal tubule (CDH1, BRN1), and podocyte (PODXL, PNA) on day 14. Scale bar, 50 μm. Values shown are the means ± SEM. P-values were determined by a Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001.