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. 2019 Dec 2;9:18091. doi: 10.1038/s41598-019-54574-4

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© The Author(s) 2019

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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Identification of BIO as inhibitor of myoblasts proliferation. (A) Schematic representation of the screening cascade: C2C12 cells were probed with the low-molecular weight LOPAC®1280 library, consisting of 1280 pharmacologically active compounds. Identification of BIO as molecule affecting cell proliferation. (B) Chemical structure of BIO compound. (C) C2C12 cells were cultured in the presence of growth medium (GM), differentiation medium (DM), BIO (3 μM) or Vehicle (DMSO), for 24 h and 48 h. The effect of BIO on both proliferation and viability of C2C12 cells was measured by CellTiter-Glo® Assay and (D) by CellTox™ Green Cytotoxicity Assay, respectively. Data were collected from three independent experiments and represent means ± SEM relative to untreated cells (according to one-way analysis of variance ANOVA using the Dunnett’s Multiple Comparison Test). (E) C2C12 cells were treated as in C and D panels and cell-cycle distribution was analyzed by flow cytometry. Data (collected from four independent experiments) represent means ± SEM of cells in each phase of the cell cycle compared to a no-treatment group.