Wire-Induced Femoral Artery Injury Model

Wire-induced injury of the femoral artery was performed at day 7 of empagliflozin treatment as previously described (19 (link)). In brief, mice were anesthetized as described earlier. A straight spring wire (0.38 mm in diameter, Cook Medical, Bloomington, IN, United States) was then inserted through the profunda femoris artery up to 1 cm into the femoral artery and left in place for 1 min to achieve an adequate wire-induced vessel injury. After removal of the wire, the profunda femoris artery was ligated (7-0 Prolene, Ethicon, Johnson & Johnson, Norderstedt, Germany) and perfusion of the dilated femoral artery was re-established. Mice were sacrificed at 10 or 21 days by cervical dislocation. The femoral artery was carefully harvested and embedded in Tissue-Tek OCT compound medium (Sakura Finetek Europe B.V., Staufen im Breisgau, Germany). Then, the arteries were snap-frozen and stored at –80°C until sectioning.