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. 2017 Jun 9;7:3209. doi: 10.1038/s41598-017-03319-2

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© The Author(s) 2017

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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APN inhibits H₂O₂-induced apoptosis. (A) C2C12 cells were treated with 5 mM H₂O₂ for the indicated periods. Samples were collected for western blotting to analyze the expression levels of Bax, Bcl–2, P53 and β-actin. (B,C) The densitometric ratios of Bax, Bcl-2 and P53 were quantified using Image J software. The analysis of the (Bax/Bcl-2)/β-actin and P53/β-actin ratios from the immunoblots is shown (data from three independent experiments). (D) The relative expression of Bax in C2C12 cells pretreated or not with 30 μg/mL APN followed by treatment with 5 mM H₂O₂ for 1 h. (E) C2C12 cells pretreated or not with various concentrations of APN followed by treatment with 5 mM H₂O₂ for 1 h. Western blotting was performed to analyze the levels of Bax, Bcl-2, P53 and β-actin. (F) Image J densitometric analysis of the (Bax/Bcl-2)/β-actin ratios from the immunoblots is shown (data from three independent experiments) (*p < 0.05, **p < 0.01).