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. 2020 Apr 29;11:2040622320916024. doi: 10.1177/2040622320916024

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© The Author(s), 2020

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).

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Figure 4. — Alkbh5 knockdown enhanced OGD/R-treated primary neuronal apoptosis and Fto overexpression attenuated OGD/R-treated primary neuronal apoptosis. (A) Primary neurons were infected with Alkbh5-shRNA and then cultured under OGD/R conditions or normoxia. Apoptosis analysis was performed and representative flow cytometry images are shown. Quantifications of the percentage of apoptotic cells are shown (mean ± SEM; n = 3; ***p < 0.001 versus Ctrl, **p < 0.01 OGD/R + Alkbh5-shRNA versus OGD/R + Ctrl-shRNA). (B) Primary neurons were infected with Alkbh5-shRNA and then cultured under OGD/R conditions or normoxia prior to performing cell activity assays. Quantifications of the percentage of primary neuronal activity are shown (mean ± SEM; n ⩾ 8; **p < 0.01 OGD/R + Alkbh5-shRNA versus OGD/R). (C) Primary neurons were infected with Alkbh5-shRNA and then cultured under OGD/R conditions or normoxia. Cell death was quantified by microscopy for PI-positive cells. Quantifications of the percentage of dead cells are shown (mean ± SEM; n = 8; *p < 0.05 versus OGD/R). (D) Primary neurons were infected with Lv-Fto and then cultured under OGD/R conditions or normoxia. Apoptosis analysis was performed and representative flow cytometry images are shown. Quantifications of the percentage of apoptotic cells are shown (mean ± SEM; n = 3; ***p < 0.001 versus Ctrl, *p < 0.05 OGD/R + Lv-Fto versus OGD/R + Lv-Ctrl). (E) Primary neurons were infected with Lv-Fto and then cultured under OGD/R conditions or normoxia prior to performing cell activity assays. Quantifications of the percentage of primary neuronal activity are shown (mean ± SEM; n ⩾ 4; **p < 0.01 OGD/R + Lv-Fto versus OGD/R). (F) Primary neurons were infected with Lv-Fto and then cultured under OGD/R conditions or normoxia. Cell death was quantified by microscopy for PI-positive cells. Quantifications of the percentage of dead cells are shown (mean ± SEM; n = 8; **p < 0. 01 versus OGD/R).

Ctrl, control; OGD/R, glucose oxygen deprivation/reoxygenation; PI, propidium iodide; SEM, standard error of the mean.