Fig 2: Agomir-212-5p suppressed the PLXNA2 and RhoA/ROCK2 in PC12 cells following OGD/R. A Representative photographs of western blots showing PLXNA2, RhoA and ROCK2 levels in each group. β-Actin was used as an internal reference. B Quantitative analysis of the western blot results. C-E Immunofluorescence staining results for the PLXNA2, RhoA and ROCK2 (in green) proteins in neurons in each group. Nuclei were stained with DAPI and are visualised in blue. Scale bar = 50 μm. F-H qRT–PCR was performed to assess the mRNA levels of PLXNA2, RhoA and ROCK2 in each group. The data are presented as the means ± SEM (n = 4–6 per group). *P < 0.05 and **P < 0.01 compared with the sham group, #P < 0.05 and ##P < 0.01 compared with the MCAO/R group

Fig 3: MiRNA sequencing of microglial exosomes 3 days after MCAO/R and qRT–PCR validation. A, B Characterisation of microglial exosomes using nanoparticle tracking analysis and transmission electron microscopy scanning. Scale bar = 200 nm. C The exosome markers CD9, CD63, and CD81 were detected using western blot analysis. D Microglia-derived exosomes were detected by western blot analysis using the microglia marker CD11b. E Heatmap showing the levels of miRNAs in microglial exosomes at 3 days after MCAO/R. F Expression of miR-30c-5p, miR-126a-5p, miR-128-3p, miR-212-5p and miR-1949 in the ischemic penumbra of the cortex at 3 days after MCAO/R was determined using qRT–PCR. G Expression levels of the miRNA-targeting genes PLXNA2, PTEN and FOXO3 in the ischemic penumbra of the cortex at 3 days after MCAO/R. H The target sites of miR-212-5p in PLXNA2 mRNA 3’ untranslated region (3’UTR). I Map of the pmirGLO luciferase reporter vector. J Dual luciferase assays revealed the binding of miR-212-5p to the 3’UTR of PLXNA2. The data are presented as the means ± SEM (n = 5 per group). *P < 0.05 and **P < 0.01 compared with the sham group. #P < 0.05 compared with the miR-NC group