Figure 4

vWF and CD31 staining in the peri-infarct lesion of MCAO mice with or without OP9-pre-conditioned UCB administration. Fluorescent immunohistochemistry was performed 3.5 months after surgery (n = 5 in the UCB + OP9 group and n = 5 in the control group, respectively). (a) Schematic illustration of the target region for the area measurement of CD31/vWF (von Willebrand factor) double-positive cells by a fluorescence microscope. One coronal section per mouse (n = 5 in each group) was analyzed. Small arteries in the brain surface of ischemic infarction (‡) and microvessels in the peri-infarct lesion located at the periphery of the ischemic infarction adjacent to the normal cortex and striatum (†) were observed. Three non-overlapping random visual fields of (†) indicated by red solid boxes (400 × magnification) per coronal section were analyzed to measure the CD31/vWF double-positive area. (b) Representative images of immunofluorescent staining of brain sections. CD31/vWF double positive cells (yellow) with CD31⁺ (green) and vWF⁺ (red) in the peri-infarct lesions (†) were subjected to the analysis in (c). Nuclei were counterstained using DAPI (blue). Scale bar: 50 μm. 200 × magnification. (c) The area measurement and cell count of CD31⁺/vWF⁺ cells per field observed in 400 × magnification (n = 15 per each group). ***p < 0.001, unpaired t-test.