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. 2020 Jul 31;16:1863–1877. doi: 10.2147/NDT.S260748

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© 2020 Bu et al.

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ATC-derived exosomal miR-361 increases OGD/R-treated PC12 cell activity. (A) OGD/R-treated PC12 cells were treated with exosomes transfected with miR-361 inhibitor or Mock at a dose of 30 μg/mL, and the expression of miR-361 in each group was detected by RT-qPCR. (B) Measurement of cell viability using MTT assay. (C) Detection of number of proliferating cells by EdU staining. (D) After Annexin V-FITC/PI labeling, the number of apoptosis was detected by flow cytometry. (E) Western blot analysis was performed to determine the contents of apoptosis-related proteins Bax, Cleaved Caspase-3 and Cleaved PARP in each group. (F) Apoptosis of PC12 cells in each group was detected by TUNEL assay. All experiments were performed three individual times; Data are expressed as mean ± standard deviation. One-way ANOVA and Tukey’s multiple comparison test were used to determine statistical significance, while in panel B and F, two-way ANOVA was used. *P < 0.05, **P < 0.01 vs the OGD/R-Mock group.

Abbreviations: ANOVA, analysis of variance; ATC, astrocyte; Bax, Bcl-2-associated X; DAPI, 4ʹ,6-diamidino-2-phenylindole; EdU, 5-ethynyl-2ʹ-deoxyuridine; Exo, exosome; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; miR, microRNA; MTT, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; OGD/R, oxygen-glucose deprivation/reoxygenation; PI, propidium iodide; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; PARP, poly (ADP-ribose) polymerase; TUNEL, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling.