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. 2018 Jul 16;8:10707. doi: 10.1038/s41598-018-29124-z

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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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AMIGO3 levels are suppressed in DRGN after injection of PEI transduced plasmids encoding shAMIGO3. (A) Low levels of AMIGO3 mRNA in IC increased significantly after DC injury (P < 0.0001) and remained high in the DC + PEI-gfp group. AMIGO3 mRNA levels reduced significantly in both DC + PEI/shAMIGO3/gfp and DC + PEI-shAMIGO3/nt3 groups (P < 0.0001). (B) Western blot and subsequent densitometry for AMIGO3 protein reflected the changes in mRNA. β-actin was used as a loading control (full scans shown). (C) Immunohistochemistry for AMIGO3 showed high levels of AMIGO3 (red) in DRGN after DC (C(i)), which were gfp⁻ (C(ii)). AMIGO3 levels remained high in DC + PEI-gfp groups (C(iv)), with gfp expression (green) in some DRGN (C(v)). AMIGO3 levels were suppressed in DRGN from DC + PEI-shAMIGO3/gfp groups (C(vii)), with gfp expression in some DRGN (C(viii)). (C(iii)), (C(vi)) and (C(ix)) are merged images showing AMIGO3 (red), gfp (green) and DAPI stained nuclei (blue). Scale bar = 100 μm. ***P < 0.0001, ANOVA.