Fig. 8.

Oral VCE-004.8 enhances ischemia-induced vessel formation and vascular endothelial cells proliferation in vivo. A Double immunofluorescence confocal staining of α-SMA (green)/CD31 (red) in gastrocnemius muscles 28 days after ischemia. Quantification of the number of vessel peer area and the perimeter of the lumen vessel. As the data cannot passed the normality test. P values are calculated using a nonparametric Kruskal–Wallis test followed by Dunn multiple. The results are shown as mean ± SEM. P values indicated in panels, significant as **p < 0.01, ***p < 0.001, ****p < 0.0001. B The number of CD31⁺/Ki67⁺ positive cells per area and number of CD31⁺/Ki67⁺ cells per total cells were quantified using double immunofluorescence confocal staining of CD31 (green) and Ki67 (red) in gastrocnemius muscles 28 day after ischemia. Significance was determined by one-way ANOVA followed by Tukey’s test. The results are shown as means ± SEM (n = 3). P values indicated in panels, significant as ****p < 0.0001. C Double immunofluorescence confocal staining of peNOS (green) and CD31 (red) in gastrocnemius muscles 10 days after ligation and quantifications. Magnified region (marked by the white rectangle) shows detailed colocalization of peNOS and CD31 expression. Scale bars equivalent to 50 μm. The data do not pass the normality test performed, and the significance was determined non-parametric followed by a Kruskal–Wallis test. Values are mean ± SEM (n = 3–4). P values indicated in panels, significant as *p < 0.1, ***p < 0.001