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. 2018 Aug 14;8:12155. doi: 10.1038/s41598-018-30615-2

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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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Mfn1 knockdown inhibits neural differentiation of iPSCs. (a–c) Cells were infected with lentiviruses containing a vector encoding shRNA directed against Mfn1 or scrambled sequence shRNA (control) for 24 h. Infected cells were selected using puromycin (1 μg/ml) for 24 h and cultured for an additional 72 h before neural differentiation. (a) The expression of OTX2 (day 2) was examined with real-time PCR. (b) The expression of PAX6 (day 4) was observed by immunocytochemistry using anti-PAX6 antibodies. Nuclei were counterstained with DAPI. PAX6 positive nuclei were counted using the ImageJ software. Bar = 100 μm. (c) The expression of Nestin (day 8) was examined with real-time PCR. Each bar represents the mean ± SD from three independent experiments. *P < 0.05.