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. 2020 Jan 31;10:1597. doi: 10.1038/s41598-020-58482-w

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© The Author(s) 2020

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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Annexin-V/7-AAD FACS analysis of PC12 cells preconditioned with the HIF PHD inhibitors followed by 24 hours re-oxygenation and 6 hours OGD. PC12 cells were exposed to PHD inhibitors (100 µM) for 24 hours in normoxia (21% O₂), followed by 24 hours re-oxygenation (21% O₂) and subsequent 6 hours OGD treatment (0.3% O₂). (A) Representative dot plots of Annexin-V/7-AAD FACS analysis of cells with the PHD inhibitor pre-treatment and 24 hours re-oxygenation followed by 6 hours OGD. Cells in the lower left quadrant represent viable cells, cells in the lower right quadrant represent early apoptosis and cells in upper right quadrant represent late apoptosis/necrosis; (B) The group data (n = 3) representing % of viable, early and late apoptotic cells. Significant reductions in the fractions of early apoptotic cells were seen in cells preconditioned with 100 µM of DMOG, FG2216, FG4592, GSK1278863 and Bay 85-3934 in comparison to 1% DMSO-treated cells following 24 hours re-oxygenation and 6 hours OGD. Data were expressed as mean ± S.D. *Indicated significance of viable cells P < 0.05 against 1% DMSO. ^#Indicated significance of apoptotic cells P < 0.05 against 1% DMSO (Two-way ANOVA, Tukey’s post-hoc analysis).