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. 2018 Aug 27;8:12875. doi: 10.1038/s41598-018-30659-4

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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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Analysis of phosphorylation of AMPK. Western blot assay was performed in the cytoplasmic fraction of the lumbar spinal cord of WT mice and VEH, LD and HD SOD1(G93A) groups (n = 3 per group). (a) Representative western blot image showing the phosphorylation rate pAMPK (62 kDa) and the expression of AMPK (62 kDa) and GAPDH (37 kDa). (b) The graph shows the densitometric analysis of the phosphorylation of pAMPK compared to the total expression of AMPK. The VEH show a slight decrease (not significant) of the phosphorylation of AMPK compare to WT. The treatment increase the pAMPK/AMPK ratio and significant differences were found between VEH and HD group (VEH vs HD, ^**P = 0.089). All gels and blots were processed in parallel showing similar results. Results were analyzed by one-way ANOVA followed by Tukey’s multiple comparisons test. Graphs are shown as mean ± SEM. ^* and ^** indicate p < 0.05 and p < 0.01 respectively of interested group vs VEH group. Full length blots are shown in Supplementary Information, in Supplemental Fig. 2.