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. 2018 Sep 19;19(9):2837. doi: 10.3390/ijms19092837

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Effect of Guanidinoacetic Acid (GAA) on proliferating myoblasts. Myoblasts were cultivated in growth medium (GM) or differentiation medium (DM) in the presence of 0, 5, 10, or 20 mM GAA. (A,B) EdU analysis showed that GAA significantly reduced the cell proliferation ratio; (C) CCK-8 tests indicated that cell growth was suppressed by GAA in a dose-dependent manner; (D) relative expression of marker genes of proliferation, including Cyclin D1 (CCND1), Cyclin Dependent Kinase 4 (CDK4) and Cyclin Dependent Kinase Inhibitor 1A (P21) after treatment with the concentration gradient of GAA; (E) images were obtained by brightfield microscopy on the fourth day of myoblast differentiation after stimulation with the concentration gradient of GAA; (F) the relative expression of myogenic differentiation 1 (MyoD). The results are expressed as mean ± S.D. of three independent experiments. * p < 0.05; ** p < 0.01 compared with the negative control (on the bars) or between the indicated groups (Tukey’s post-hoc test).