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. 2020 Sep 2;14:559384. doi: 10.3389/fncel.2020.559384

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Copyright © 2020 Si, Li, Huang, Ye, Li, Li, Kuang, Chen and Zhu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The apoptosis level and percentage of live cells at different time points of reperfusion under OGD/R injury condition in PC12 cells. (A) Apoptosis levels in PC12 cells at 0, 6, 12, 18, and 24 h of reperfusion after 6 h of OGD stimulation. Annexin V-FITC/PI double staining assay by the flow cytometry was used to detect the dynamic change of apoptosis levels. A total of 10,000 cells were counted for each flow cytometry analysis. ****, compared with R-0 h group; P < 0.0001. X-axis: Annexin V; Y-axis: PI. (B) BCL2 and Caspase 3 active protein expression at different time points of reperfusion. The western blotting assay was used to detect the protein expression BCL2 and Caspase 3 active at different time points (0, 6, 12, 18, and 24 h) of reperfusion after 6 h of OGD stimulation. ****, compared with R-0 h group; P < 0.0001. (C) The percentage of live cells at different time points of reperfusion under OGD/R injury condition. Live/Dead cell staining was used to measure the percentage of live cells at different time points of reperfusion. Cells were stained with Calcein AM (green; live cells) and PI (red; dead cells). ****, compared with R-0 h group; P < 0.0001. Scale bars: 25 μm.