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. 2018 Feb 9;8:2710. doi: 10.1038/s41598-018-21182-7

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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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Effect of higher doses of cocaine on ROS generation and membrane integrity. Differentiated cells in phenol-red free medium were treated with PBS (control) or cocaine for 48 h. ROS generation in the cells loaded with DCFDA probe (10 μM) was measured (n = 12, **P = 0.0003) with the excitation and emission filters set at 485 nm and 530 nm respectively in a fluorescence micro plate reader (A); membrane integrity was determined by measuring LDH release (B) from the cells (n = 10, **P = 0.001) at 490 nm in a micro plate reader. Data are expressed as mean ± SEM, significant compared to the PBS controls.