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. 2020 Mar 27;36(7):733–760. doi: 10.1007/s12264-020-00480-z

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Protective effect of hypoxic preconditioning and BDNF overexpression 24 h after OGD. A Double staining of cells with Hoechst 33342 (HO342) and propidium iodide (PI) and merged images (Merge) 24 h after 40-min OGD. B Cytogram of the viability of hippocampal cells (X-axis, intensity of PI fluorescence; Y-axis, intensity of Hoechst 33342 fluorescence; cells were stained with probes 24 h after HP). C Effects of HP, BDNF, and its knockdown on the induction of necrosis and apoptosis 24 h after OGD. Percentages of living cells (black) and cells in which early apoptosis (violet), apoptosis (green), and necrosis (red) were detected. Cultures were returned to a CO₂ incubator for 24 h after HP, then used in OGD experiments (40-min OGD) and returned to the CO₂ incubator for 24 h, after which, they were stained with HO342 and PI. (***P ≤ 0.001, HP vs Scrambled, AAV-BDNF + HP vs Scrambled, and BDNF-KD + HP vs Scrambled; **P = 0.008, HP vs AAV-BDNF + HP; ***P ≤ 0.001, HP vs BDNF-KD + HP and AAV-BDNF + HP vs BDNF-KD + HP).