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. 2018 Mar 23;8:5101. doi: 10.1038/s41598-018-23293-7

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© The Author(s) 2018

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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In vitro differentiation of P19 cells mimics early processes in mammalian neuroectodermal development in vivo. (a) and (b) Confocal images of P19 cells stained with anti-Oct4 and anti-NANOG antibody (FITC-green), respectively. (c) Confocal images of P19-derived neural progenitor cells (NPCs) on the second day of neuronal induction upon plating on adhesive tissue culture dishes stained with anti-Pax6 (FITC-green) and anti-nestin (FITC-Texas). (d) Neuronal precursors become post-mitotic and undergo end-terminal differentiation, observed by confocal microscopy of neurons stained with anti-Tubb3 on day 6 of neuronal differentiation upon plating on adhesive tissue culture dishes. Cell nuclei were counterstained with DAPI. Images of immuno-staining and DAPI staining were overlaid. Scale bar represents 10 micrometers.