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. 2019 Oct 31;9:15717. doi: 10.1038/s41598-019-52083-y

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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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Effect of AC on mitochondrial oxidative stress in NSCs injured by OGD. (A) NSCs were treated with OGD for 30 m and free radical generation was measured using the fluorescent probe DCFH-DA. OGD significantly increased levels of intracellular reactive oxygen species (ROS). However, co-treatment with AC dramatically decreased ROS levels in NSCs. (B) Measurement of intracellular Ca²⁺ levels suggested that intracellular Ca²⁺ influx into NSCs was increased by OGD but is decreased by AC treatment. (C) Mitochondrial membrane potential was measured in NSCs under diverse conditions and it decreased significantly upon OGD treatment, whereas AC effectively restored membrane potential to almost normal. (D) ATP level decreased with OGD while AC increased ATP level. (E) Western blots show that expression of proteins associated with cell survival, such as Bcl-2 and mitochondrial cytochrome c, decreased in response to OGD treatment but increased with AC treatment. Proteins related to cell death, including Bax, cytosolic cytochrome c, and cleaved PARP increased with OGD treatment but decreased with AC treatment. Data are means (% of control) ± SD from five independent experiments. Treatment groups were compared with the control group using Tukey’s test after one-way ANOVA. *p < 0.05 and **p < 0.01 (vs. control group), ^#p < 0.05 (vs. the group treated only with OGD). Full-length blots/gels are presented in Supplementary Fig. S6.