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. 2019 Aug 16;9:11945. doi: 10.1038/s41598-019-48421-9

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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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HD-PTP expression in embryonic motor neurons and CRISPR-mediated depletion. (a) Representative images of chick embryonic spinal cord sections at HH st. 25 and HH st. 28 where ISL1 and PTPN23 (chicken HD-PTP-encoding gene) mRNA was detected using in situ hybridisation. Note expression of PTPN23 in ISL1-expressing motor column (arrows). (b) Representative images of anti-HD-PTP antibody staining in growth cones and cell bodies of dissociated motor neurons harvested from embryonic spinal cords electroporated with Control^CRISPR or HD-PTP^CRISPR plasmids. (c) Quantification of HD-PTP signals in growth cones of dissociated motor neurons harvested from embryonic spinal cords shows a decreased signal in HD-PTP^CRISPR compared to Control^CRISPR (n = 3, 10–12 growth cones/n; p = 0.0023; Student’s t-test). (d) Quantification of HD-PTP signal in cell bodies of dissociated motor neurons harvested from embryonic spinal cords show decrease signal in HD-PTP^CRISPR compared to Control^CRISPR (n = 3, 30–50 cell bodies/n; p = 0.0009; Student’s t-test). Values are plotted as mean ± SD. All values can be found in Supplementary Table S4. ***p < 0.001; **p < 0.01. Visible light images (a) and inverted grayscale fluorescent images (b).