Effects of the Insulted Neuronal Cells-Derived Extracellular Vesicles on the Survival of Umbilical Cord-Derived Mesenchymal Stem Cells following Cerebral Ischemia/Reperfusion Injury

Figure 7

Impact of silencing Rab27a on the release of EVs from N2A cells and injury to UC-MSCs. N2A cells were transfected with Rab27a siRNA. These cells were verified with western blotting, mRNA and EVs markers. (a–c) Silencing of Rab27a on N2A cells as evaluated by western blotting and mRNA. (d) Western blotting analysis of EVs markers in EVs and whole cellular lysis (WCL). (e) The concentration of EVs as evaluated by NTA. Next, UC-MSCs were cultured with medium from Rab27a-siRNA-CMs group and Vector-N2A-CMs group under non-OGD/R condition and OGD/R24H insult. (f–g) Apoptosis of UC-MSCs in Rab27a-siRNA-CMs group and Vector-N2A-CMs group mediums under non-OGD/R condition and OGD/R24H insult were detected by flow cytometry assay. (h) Apoptosis of UC-MSCs as detected by LDH leakage assay. (i) Cell viability of UC-MSCs as determined by MTT assay. (j–k) Expression and quantitative data of cleaved-caspase3, cleaved-caspase9, and cytochrome C were determined by western blotting. (l) Levels of ROS as determined by dihydroethidium-DCFH-DA assay. (m) Levels of ATP as determined by firefly luciferase assay. (n) Levels of SOD as determined by WST-8 assay. (O) Levels of T-AOC as detected by ABTS assay. All data are presented as the (). ; , control group compared with Rab27s siRNA group. Rab27a-siRNA-N2A-CMs group compared with Vector-N2A-CMs group.

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