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Research Paper Volume 13, Issue 3 pp 4079—4095

Microglia exosomal miRNA-137 attenuates ischemic brain injury through targeting Notch1

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Figure 7. M2-phenotype microglia-derived exosomes (BV2-Exo) attenuated tMCAO-induced neuronal apoptosis through Notch1. (A) Protein expression of Notch1 in brains of mice treated with 1) control, 2) tMCAO plus BV2-Exo, 3) tMCAO plus BV2-Exo+miRNA-137-IN, 4) tMCAO plus BV2-Exo+miRNA-137-IN and Crenigacestat, and 5) tMCAO plus BV2-Exo+miRNA-137-NC, as detected by Western blotting assay. (B) Modified neurological severity score for mice treated with 1) control, 2) tMCAO plus BV2-Exo, 3) tMCAO plus BV2-Exo+miRNA-137-IN, 4) tMCAO plus BV2-Exo+miRNA-137-IN and Crenigacestat, and 5) tMCAO plus BV2-Exo+miRNA-137-NC. (C) Relative infarct volume in brains of mice treated with 1) control, 2) tMCAO plus BV2-Exo, 3) tMCAO plus BV2-Exo+miRNA-137-IN, 4) tMCAO plus BV2-Exo+miRNA-137-IN and Crenigacestat, and 5) tMCAO plus BV2-Exo+miRNA-137-NC, displayed as brain cresyl violet staining and brain tissues of ischemic mice treated with indicated treatments. Yellow dotted boxes represent the infarct areas. (D) Double-staining of NeuN/TUNEL in brain sections of mice treated with 1) control, 2) tMCAO plus BV2-Exo, 3) tMCAO plus BV2-Exo+miRNA-137-IN, 4) tMCAO plus BV2-Exo+miRNA-137-IN and Crenigacestat, and 5) tMCAO plus BV2-Exo+miRNA-137-NC. Red color indicated NeuN and green color indicated TUNEL staining. Scale bar = 25 μm. (E) Protein expression of caspase-3 and cleaved caspase-3 in brains of mice treated with 1) control, 2) tMCAO plus BV2-Exo, 3) tMCAO plus BV2-Exo+miRNA-137-IN, 4) tMCAO plus BV2-Exo+miRNA-137-IN and Crenigacestat, and 5) tMCAO plus BV2-Exo+miRNA-137-NC, as detected by Western blotting assay. Data are presented as mean±SD. *, p<0.05. At least three replicates were available for statistical analysis in each treatment.