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. 2018 Apr 4;8:5646. doi: 10.1038/s41598-018-24017-7

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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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In-gel activity assays for electron transport chain (ETC) complexes from adult mice. (a) Representative images of complexes I-IV and ATP synthase of the ETC, assayed following blue native polyacrylamide gel electrophoresis of liver mitochondrial homogenates to demonstrate the effect of metformin in injured adult animals. (b) Densitometric measurements of in-gel activity assay bands using ImageJ for Windows. (c) Bands selected from the gel following staining with Coomassie Blue G-250 served as a loading control. In-gel activity assays from panel a were performed in the absence of key reaction mixture substrates (d) or in the presence of inhibitors (e) to confirm band specificity. (f) Respiration profiles of liver mitochondria from adult control mice (grey; n = 9) and control mice given metformin (green; n = 9). : control (n = 5); ■: burn (n = 5); : burn + metformin treatment (n = 5). Values are presented as mean ± standard deviation. *p ≤ 0.05; **p ≤ 0.01.

Inline graphic — In-gel activity assays for electron transport chain (ETC) complexes from adult mice. (a) Representative images of complexes I-IV and ATP synthase of the ETC, assayed following blue native polyacrylamide gel electrophoresis of liver mitochondrial homogenates to demonstrate the effect of metformin in injured adult animals. (b) Densitometric measurements of in-gel activity assay bands using ImageJ for Windows. (c) Bands selected from the gel following staining with Coomassie Blue G-250 served as a loading control. In-gel activity assays from panel a were performed in the absence of key reaction mixture substrates (d) or in the presence of inhibitors (e) to confirm band specificity. (f) Respiration profiles of liver mitochondria from adult control mice (grey; n = 9) and control mice given metformin (green; n = 9). : control (n = 5); ■: burn (n = 5); : burn + metformin treatment (n = 5). Values are presented as mean ± standard deviation. *p ≤ 0.05; **p ≤ 0.01.