Figure 7

Evaluation of the hypEV effects on brain angiogenesis in a mouse model for ischemic stroke. After MCAO, mice were injected intraperitoneally with PBS or hypEVs, immediately after the procedure and each 24 h, for a total of 5 consecutive days. Mice were perfused at day 7 post-MCAO (n = 5 PBS, n = 6 hypEVs). (A) Representative pictures of blood vessels in the perilesional area. Immunofluorescence on brain sections was performed using anti-CD31 antibodies (endothelial cells) and Hoechst (nuclei). Images were acquired at a confocal microscope, using a 25× objective. Scale bar: 50 µm. (B) Quantification of the CD31 signal in the perilesional region was performed using ImageJ, fixing a threshold to detect the percentage of the CD31⁺ area, expressed as arbitrary units (a.u.), for each field of view analysed. (C) Analysis of proliferating nuclei in the perilesional area. The number of proliferating cells per field of view analysed is reported. Quantification of the Ki67⁺ area was performed using ImageJ and is expressed as normalised to controls. Bars represent mean ± SEM. The non-parametric t-test was applied. * p < 0.05.