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. 2019 Jun 21;9:9013. doi: 10.1038/s41598-019-45247-3

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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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Comparison of in vivo differentiation of Zfp488-ZsGreen1-hNSCs and control-hNSCs into astrocytes within the demyelinating lesion in the CC using GFAP antibody (A,D, low power). Co-labeled GFAP⁺ (red) (B1), and ZsGreen1⁺ cells (B2) revealed that only a few double-labeled cells (B, arrows) among the transplanted surviving Zfp488-ZsGreen1-hNSCs had differentiated into an astrocytic cell lineage. In contrast, numerous transplanted control ZsGreen1-hNSCs were found to differentiate into GFAP⁺ astrocytes (red, fig. C, arrows), as shown by co-labeling of GFAP (red, C1), and ZsGreen1 reporter (C2). Quantitative analysis of the confocal images showed that 9.8 ± 2.6% in Zfp488-hNSCs differentiated into GFAP astrocytes while 42.0 ± 2.2% in control-hNSCs differentiated into GFAP⁺ astrocytes (see bar graph). Blue nuclei, Hoechst staining. Bar (for all panels) is equal to 100 μM. Abbreviations: CC, corpus callosum; GFAP, glial fibrillary acidic protein; ZsGreen1, Zoanthus sp green fluorescent protein.