Mouse model of atherogenic remodeling

From six weeks of age, mice were on continuous western diet (Ssniff^® TD88137, 0,3% cholesterol) for a total of 10 weeks. At 7 weeks of age Cre recombinase was activated in EC-PRAS40-KO mice by intraperitoneal injections of tamoxifen (Sigma, T5648, 1 mg/animal/day on five consecutive days). Cre-negative littermate control mice underwent the same procedure.

At 12 weeks of age partial carotid ligation was performed as described by us before²⁸. Briefly, mice were anaesthetized by intraperitoneal injection of ketamine (120 mg/kg, Pfizer, Germany) and xylazine (16 mg/kg, Bayer; Germany) and placed on a heated surgical pad. After hair removal a midline cervical incision was made and the distal branches of the left common carotid artery were exposed. Three of the four branches (external carotid artery, internal carotid artery and occipital artery) were then ligated distal to the superior thyroid artery branch off, which was left open as an outflow (see schematic diagram in Fig. 6a). 6.0 silk sutures (Serag-Wiessner) were used for ligations; the skin was sutured with absorbable 6.0 silk suture (CatGut, Germany). After surgery, animals were monitored in a chamber on a heating pad until recovery. Western diet was continued until harvesting of carotid arteries 4 weeks after partial carotid ligation.

All animal experiments were approved by the animal ethics committee of the federal state of Berlin (Tierversuchskommission des Landes Berlin, Germany) and were performed accordingly under adherence to the effective guidelines and the German Protection of Animals Act.