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. 2018 Oct 23;8:15647. doi: 10.1038/s41598-018-34092-5

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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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Knockdown of Plexin-A1 induces the formation of supernumerary axons. (a,b) Hippocampal neurons from E18 rat embryos were transfected with vectors for GFP (green) and shRNAs against Plexin-A1, -A2. -A3, or -A4 or pSUPER (control) and analyzed at 3 d.i.v. by staining with an anti-MAP2 (red, dendrites) and the Tau-1 antibody (blue, axon). Representative images of transfected neurons are shown. The scale bar is 25 μm. (b) The percentage of unpolarized neurons without an axon (0, red), polarized neurons with a single axon (1, blue), neurons with multiple axons (>1, green) and neurons with neurites that are positive for both axonal and dendritic markers (in, yellow) is shown (Student’s t-test and two-way ANOVA; n = 3, independent experiments with >150 neurons per experiment; values are means ± s.e.m, **p < 0.01 compared to control). (c) Hippocampal neurons from E18 rat embryos were transfected with vectors for GFP (green) and an shRNA against Plexin-A1 (RNAi) or pSUPER (control) and analyzed at 3 d.i.v. by staining with an anti-Plexin-A1 (red) and the Tau-1 antibody (blue, axon). Representative images of transfected neurons are shown. The scale bar is 25 μm.