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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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Muscle-specific genes related to hypertrophy regulated by EE. Differentiated C2C12 cells were treated with serum free medium (CTL), EE (100 μg/ml), IGF-1 (20 ng/ml), or testosterone (T, 500 nM). The change of indicated gene expression after 24 h treatment with aforementioned regimens was assessed by qPCR or western blot. (A) While IGF-1 significantly down-regulated the expression levels of MRF4, MAFbx and MuRF1 genes, EE could only down-regulate MRF4; and T has no regulatory effect (N = 5). (B,C) The expression of myostatin (MSTN; full-length precursor, 43 kDa) was down-regulated by IGF-1, EE and T (N = 4). The abundance of the PCR amplified fragment of the interested gene was measured and expressed as 2^−ΔΔCT after a correction to the simultaneously amplified GAPDH. All data were expressed as mean ± SD. The symbol * stands for p < 0.05 as compared to basal (CTL); the symbol ^# stands for p < 0.05 as compared to T.