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. 2018 Jul 12;8:10579. doi: 10.1038/s41598-018-28605-5

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© The Author(s) 2018

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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Suppression of osteoblastic differentiation of C2C12 cells by introduction of Mct1 siRNA or inhibition of MCT-1 activity. C2C12 cells introduced with control or Mct1 siRNA were cultured for 72 (a,b) or 48 (c) hours in the absence (−) or presence (+) of BMP-2 (300 ng/mL). (a) Results of ALP activity staining. (b) ALP activity in lysates of C2C12 cells. (c) Expressions of mRNAs for Tnap, Runx2, and Sp7 were determined using real-time RT-PCR, then normalized against that of Gapdh, with the results shown as relative values. (d) C2C12 cells were cultured for 48 hours in the presence or absence of 1 μmol/L of AZD3965, a specific inhibitor of MCT-1, and with (+) or without (−) BMP-2 (300 ng/mL). ALP activity was detected by staining. (e) ALP activity in lysates of C2C12 cells cultured under the same conditions. (b,c,e) Values are expressed as the mean ± SD (n = 3–5). **Significantly different from control group (p < 0.01).