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. 2017 Oct 10;7:12931. doi: 10.1038/s41598-017-12788-4

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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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6E11 protects HAEC from cold hypoxia reoxygenation injury when treatment occurs during cold hypoxia step (a), during both cold hypoxia and reoxygenation steps (b), but not when treatment occurs during the reoxygenation step (c). Increasing concentrations (1, 3.33, 10, 33.3 µM) of 6E11, Nec-1, Nec-1s or 8A03 were added to the UW preservation solution during hypoxia or/and in PBS – 2% FBS during reoxygenation. UW corresponds to cells treated with UW preservation solution. Controls are cells not subjected to this protocol and continuously oxygenated. DMSO are cells treated with drug vehicle. The ratio of cell viability evaluated by XTT test, is calculated in comparison to the untreated control (100% of cell viability). Data shown are mean ± SD, n = 3; ANOVA and Tukey’s Multiple Comparison Test; *P < 0.05 vs UW. See Supplementary Fig. S3 for a schematic representation of the protocol.