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. 2019 Feb 21;9:2516. doi: 10.1038/s41598-019-38676-7

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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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Afatinib attenuated OGD-induced increases in COX-II, iNOS and NO levels. (A) CTX-TNA2 cells were exposed to oxygen-glucose deprivation (OGD) for various durations (3, 6, 12 h). Western blot assay was employed to measure COX-II, iNOS and β-Actin. Each lane contained 40 μg protein for all experiments. Graphs show statistical results of COX-II and iNOS from relative optical density of bands on the blots. Values are the mean ± S.E.M. (n = 3/group).*P < 0.05 in the OGD group compared with the control by t-test. (B) CTX-TNA2 cells were exposed to 12-h OGD. Afatinib (1, 10 nM) was included in the culture medium concomitantly with OGD exposure. Graphs show statistical results of COX-II and iNOS levels from relative optical density of bands on the blots. Values are the mean ± S.E.M. (n = 3/group). (C) CTX-TNA2 cells and (D) Primary cultured astrocytes were exposed to 24 h-OGD or 12 h-OGD, respectively. Afatinib (1, 10 nM) was included in the culture medium concomitantly with OGD exposure. The levels of NO were measured using Griess reaction. Values are the mean ± S.E.M. (n = 3/group). *P < 0.05 in the OGD group compared with the control, ^#P < 0.05 in OGD plus afatinib compared with OGD alone by Kruskal-Wallis test and followed by Mann-Whitney U test as post-hoc method.