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. 2019 Jul 23;9:10646. doi: 10.1038/s41598-019-47123-6

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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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Use of nicotinamide, Activin A and ChiR99021 in a sequential manner recapitulates the main steps of retinal development. (A) Schematic representation of the retinal development. H, Hypothalamus; OV, Optic Vesicle; L, Lens; NR, Neural Retina; RPE, Retinal Pigment Epithelium; OS, Optic Stalk. (B) Relative gene expressions were quantified by RT-qPCR and normalized to mRNA expression at day 0 (n = 3, mean ± SD). Control condition corresponds to RPE 20% KSR medium. (C) Representative immunofluorescence for PAX6 and LHX2 at day 7 and for VSX2 and MITF at day 10 (D), D14 (E) and D21 (F). Nuclei are stained with DAPI (blue).