(A) A group of 8- to 12-week-old C57BL/6 female mice were initiated on probenecid (n = 7) or probenecid plus IS (n = 7) and underwent HLI followed by harvest after 5 days. (B) Representative images of the posterior calf muscles from the ligated limbs of mice stained for α-actin and CD31. Total of 30 images per group (n = 7 mice/group). Insets show myocyte with surrounding capillaries. Scale bars: 25 μm. Original magnification ×400. (C) Integrated density of CD31⁺ was normalized to that of α-actin and presented as a ratio. A total of 30 images from 7 mice/group are represented, and the line represents the median. Student’s t test was performed. ***P = 0.001. (D) Lysates of posterior calf muscles of the ligated limb of mice were probed. Representative immunoblots from 3 separate mice from each group (n = 7 mice/group). (E) Normalized β-catenin in muscle lysates is presented in box-and-whisker plot. Student’s t test was performed. **P = 0.007. (F) Four to five random images of posterior calf muscles from the ligated limbs of mice were stained for β-catenin and CD31. Insets show a myocyte with β-catenin with surrounding capillaries. Blue dotted line represents ROI of a muscle surrounded by capillaries, and white dotted line represents the ROI of a muscle. White asterisk corresponds to β-catenin in a muscle. White arrowhead is directed to β-catenin in the capillary. Scale bars: 25 μm. Original magnification, ×400. (G) First, the integrated density was estimated from the ROI of area surrounded by blue dotted line (β-catenin in a muscle with surrounding capillary). Next, the integrated density was analyzed only from area surrounded by white dotted line (β-catenin in muscles). Their differences correspond to β-catenin in the capillaries. The integrated densities of capillaries and muscles from 30 random images (n = 7 mice/group) are shown. Lines represent the median. Student’s t test was performed. ***P < 0.001; *P = 0.05.