Hindlimb Ischemia in Dystrophic Mice

Procedures involving living animals were approved by local ethics committees and were performed according to the Guidelines of the Italian National Institutes of Health (Art. 31 D.lgs 26/2014, 4 March 2014). Animals used in the study were 3-month-old dystrophic C57BL/ 10ScSn Dmd^mdx and age-matched wild-type control mice provided by Charles River (Calco, Lecco, Italy). Postoperatively, animals were administered by intraperitoneal injection of the clinically approved immunosuppressive drug tacrolimus (FK-506; Sigma-Aldrich St. Louis, MO, USA) 2 mg/kg per day [32 (link)]. Acute hindlimb ischemia was induced by removal of the femoral artery, as described previously [33 (link)]. Measure of the blood flow in the ischemic hindlimb compared to the contralateral control was performed by laser Doppler perfusion imaging (Lisca Inc., North Brunswick, NJ, USA).