Representative inverted grey scale images of iso-lectin staining (A) were used to calculate capillary to muscle fiber ratio (n = 4 per condition) (B). In (C), VEGF protein levels were assessed by Western blotting with expression normalized to ß-actin (n = 3 for sham + vehicle, n = 4 for overload + vehicle, overload + losartan, and for sham + losartan). Skeletal muscle endothelial cells (n = 4 per condition) (D) were treated overnight with the AT1R inhibitor Losartan (0.1 or 1 µM) and in (E) cells were treated with Ang II (0.1 µM) or Ang II and losartan (1 µM) overnight. Cells were lysed for qPCR analysis of VEGF mRNA levels. Values are presented as mean ± SEM. There was a significant main effect of overload on capillary to muscle fiber ratio (*), as assessed by two way ANOVA. For VEGF protein levels, two way ANOVA revealed a significant main effect of overload (*) and the losartan treatment (**) as well as a significant difference between overload and overload + losartan conditions and the losartan and overload + losartan conditions (#) by Bonferroni’s multiple comparison test. In (D), *denotes p<0.05 vs. untreated cells and in E, *denotes p<0.05 vs. Ang II treatment alone. S-sham, O-overload and L-losartan.