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. 2022 Jan 17;12:827. doi: 10.1038/s41598-022-04804-z

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© The Author(s) 2022

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 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/.

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(A) Representative images of phase-contrast and immunostaining of MHC I and II in C2C12 cells at day 7 in Serum, B27, and AIM-V cultures. Images show immunostaining for MHC I (green), MHC II (red) and Hoechst counterstained nuclei (blue). Merged panels show composite images. Scale bar indicates 100 µm. Quantification of MHC I and II images from A was shown following figures. The positively MHC I and II—stained area was normalized to nuclei area. (B) Quantification results of MHC I (slow type)—positive stained myotubes. (C) Quantification of MHC II (fast type)-positive stained myotubes. (D) The ratio of quantified MHC I to MHC II, indicating slow type of muscle is more dominant. The experiments were performed three times and representative results are shown. (E) The sum of the glycolytic metabolism-related metabolites (G6P, F6P, F1, 6BP, 2/3-PG, PEP, Pyr, and Lac) and (F) Oxidative metabolism-related metabolites (Cit, Icit, aKG, Suc, Fum, IA, and Mal). (G) The ratio of sum of oxidative to glycolytic-related metabolites. Data presented as average ± standard deviation (SD). (n) = 4. *p < 0.05, **p < 0.01, ***p < 0.001.