Fig. 7

Neuroprotection of EXO-Hep in tMCAO rats. A After purified astrocytic mitochondria were labelled with Mito Tracker Red and injected into the cortex on the ischemic side of the rat, the colocalization of red labelled astrocytic mitochondria with neurons labelled with anti-NeuN (green) was observed by immunofluorescence staining. Scale scale is 50 μm. B Representative immunofluorescence staining and quantitative analysis for NeuN positive cells (green) after treated with purified mitochondria from A1-AS treated with Hep, EXO and EXO-Hep in ischemic cerebral tissue of tMCAO rats at 2 h of ischemia following by 24 h reperfusion. NeuN antibodies were used to stain neurons in ischemic cerebral tissue of tMCAO rats. DAPI (blue) was used as a nuclear marker. Quantification of the number of NeuN positive neurons were presented in panel. The relative intensity of NeuN was determined by calculating the ratio of fluorescent intensity of NeuN in groups to fluorescent intensity of NeuN in sham group. The scale bar in all Fig. 7B is 100 μm. Data are expressed as means ± SD (n = 3), ***P < 0.001. C Representative brain slices with infarcts were stained by 2,3,5-triphenyltetrazolium chloride (TTC) and infarct volume was calculated in tMCAO rats treated with purified mitochondria from A1-AS treated with Hep, EXO and EXO-Hep at 2 h of ischemia following by 24 h reperfusion. Data are expressed as means ± SD (n = 3), ***P < 0.001. In addition, Zea-Longa neurological scores and Ludmila Belayev neurological scores were determined. Data represent means ± SD (n = 3),*P < 0.05, **P < 0.01. D Representative brain slices with infarcts were stained by 2,3,5-triphenyltetrazolium chloride (TTC) and infarct volume was calculated in tMCAO rats treated with Hep, EXO and EXO-Hep at 2 h of ischemia following by 24 h reperfusion. Data are expressed as means ± SD (n = 3). ***P < 0.001. In addition, Zea-Longa neurological scores and Ludmila Belayev neurological scores were determined. Data represent means ± SD (n = 3), *P < 0.05, **P < 0.01. E Representative immunofluorescence staining and quantitative analysis for NeuN positive cells (green) in ischemic cerebral tissue of tMCAO rats treated with Hep, EXO and EXO-Hep at 2 h of ischemia following by 24 h reperfusion. NeuN antibodies were used to stain neurons in ischemic cerebral tissue of rats. DAPI (blue) was used as a nuclear marker. The relative intensity of NeuN was determined by calculating the ratio of fluorescent intensity of NeuN in groups to fluorescent intensity of NeuN in sham group. The scale bar in all Fig. 7E is 100 μm. Data are expressed as means ± SD (n = 3), ***P < 0.001. F Representative immunofluorescence staining and quantitative analysis for C3 positive cells (red) in ischemic cerebral tissue of tMCAO rats treated with Hep, EXO and EXO-Hep at 2 h of ischemia following by 24 h reperfusion. C3 antibodies were used to stain type A1 astrocytes in ischemic cerebral tissue of tMCAO rats. DAPI (blue) was used as a nuclear marker. The relative intensity of C3 was determined by calculating the ratio of fluorescent intensity of C3 in groups to fluorescent intensity of C3 in sham group. The scale bar in all Fig. 7F is 50 μm. Data are expressed as means ± SD (n = 3), **P < 0.01, ***P < 0.001