Fig. 3

Arteriogenesis in hindlimb ischemia after ADSCs delivery. A, D Representative images of arterioles—double immunostaining of endothelial cells (CD31, green) and arterioles (αSMA, red) in: healthy, ischemic, intramuscular, intravenous and contralateral groups on the 7th and the 14th days post-injury (dpi). Nuclei stained with DAPI (blue), scale bars = 50 μm. B, E The number of arterioles (αSMA⁺ cells) was counted in collected quadriceps (B) and gastrocnemius (E) muscles on the 7th and the 14th dpi. The number of arterioles in each group was determined in 5 fields of view per animal (n = 3–5) at a magnification of 20× and calculated per 1mm² of tissue. Each column in the graph represents a mean ± SE. C, F The average arteriole diameter was counted in quadriceps (C) and gastrocnemius (F) muscles collected on the 7th and the 14th dpi. The average arteriole diameter was calculated using ImageJ software in 5 fields of view per animal (n = 3–5) at a magnification of 20×. Results are presented as mean ± standard error (SE) or in box plot as median, quartiles and min/max values as whiskers. #p < 0.05, ##p < 0.005 by the ANOVA followed by the Tukey’s post hoc test. *p < 0.02, **p < 0.002, ***p < 0.0001 evaluated with Kruskal–Wallis one-way analysis of variance and multiple comparison of mean ranks for all groups