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. 2017 Sep 20;7:12028. doi: 10.1038/s41598-017-11086-3

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© The Author(s) 2017

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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Molecular markers of regeneration are modulated by NH treatment. (A) Histogram showing the average number (±SEM) of MNs labeled after intramuscular injection of True Blue into the TA and GA on the injured side. The number of positive MNs were counted in the L4-L5 segments of the spinal cords from animals at 6 months post RE for animals in RE group (not treated) or NH-treated groups (RE + NH). (B) Left, Representative microphotographs of GAP43 immunolabeling of neuronal processes at the white matter ventral horns from injured animals. Right, Bar graph of the mean immunolabeling intensity for GAP43 in a region inside the white matter from injured spinal cords. (C,D) Left, Confocal images of MNs immunolabeled for (C) phosphorylated AKT at S-473 and D) p70S6K at T-389 and counterstained with Fluoro Nissl Green and DAPI (blue) in injured spinal cord at 6 months post RE. Right, Histograms of the mean of the immunofluorescence intensity for each marker inside the cytoplasm of injured MNs (n = 4 animals, t-test, *p < 0.05 RE + NH vs. RE). Scale bar = 100 µm in B; 25 µm in C and D.