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. 2021 May 24;11:10790. doi: 10.1038/s41598-021-89039-0

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© The Author(s) 2021

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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Effects of EE and ICA on AMPK/Thr172 phosphorylation. Differentiated C2C12 cells were starved in serum-free medium overnight before EE (100, 200 μg/ml), ICA (1–5 μM), and IGF-1 (20 ng/ml) treatment. Representative western blot images and corresponding densitometry measurements were shown and presented as phosphorylated/total protein ratios. (A) A time interval ranging from 15–120 min showed a sequential appearance of the phosphorylated AMPK induced by EE (100 μg/mL) (N = 5). (B) After a 2-h treatment, AMPK was activated by EE, but not by IGF-1 (N = 3). (C) ICA (1–2 μM) treatment induced a similar degree of AMPK phosphorylation as EE did (N = 3). All data were expressed as mean ± SD. The symbol * stands for p < 0.05 as compared to CTL, the symbol ^# represents p < 0.05 as compared to IGF-1.