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. 2020 Oct 22;10:30. doi: 10.1186/s13395-020-00249-y

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Metabolomic signatures of NAD replacement strategies in MDX muscle. a Heatmap of ion counts for polar metabolites reaching significance (adjusted p < 0.05) in MDX muscle compared to WT controls (left column) aligned with the same metabolites identified in GSK978A-treated (center column) and NR-treated (right column) muscles after 20 weeks of treatment. b Correlation of the changes in ion abundance induced by GSK978A and (c) NR supplementation in MDX muscle after 20 weeks compared to changes driven by genotype alone. Colored points indicate metabolite ions significantly altered in at least one of the comparisons and the dashed line indicates the least-squares regression of these points. R indicates the correlation coefficient of the significant (adjusted p < 0.05) ions and p indicates the significance level of the associated Pearson’s correlation. d Volcano plot of biochemical pathways significantly altered in MDX muscle by GSK978A. Significantly altered metabolites were used to calculate pathway impact and highly affected pathways are indicated. e Total NAD content of gastrocnemius muscles harvested from contralateral limbs. Significance was determined by one-way ANOVA with Tukey’s post hoc test (**p < 0.01; ns, not significant). f Box plots of ion abundances for representative members of glycolysis (G6P, F6BP, G3P), pentose phosphate pathway (R5P), and TCA cycle (CA, SA). N = 9-12 mice per group. Whiskers represent minimum and maximum values. Significance was determined by one-way ANOVA with Tukey’s post hoc test (p values adjusted for multiple testing)