Heparinized polyethylene tubes

In both experiments, at day 20 of lactation, blood from Vena jugularis was collected 3 h after feed intake into heparinized polyethylene tubes (Sarstedt, Nürnberg, Germany), and plasma was obtained by centrifugation of the blood (1100 × g, 10 min, 4°C) and stored at -20°C pending analysis. At the same day, liver and skeletal muscle biopsy samples were taken percutaneously after anaesthesia by intravenous injection with 2 mg azaperon (Stresnil, Janssen-Cilag GmbH, Neuss, Germany) per kg body mass, 20 mg ketamine (Ursotamin, Serumwerke Bernburg AG, Germany) per kg body mass and up to 2.4 mg thiopental (Thiopental Inresa 0.5 g, Freiburg, Germany) per kg body mass as required for maintenance of anaesthesia. The biopsy procedure has been described recently in detail [36 (link)]. Liver samples were immediately snap-frozen and stored at -80°C pending analysis.

At day 29, the pigs of both experiments were killed in the post-prandial period (2 h after their last meal) by electronarcosis followed by exsanguination. Blood was collected into heparinized polyethylene tubes (Sarstedt, Nümbrecht, Germany) and plasma obtained by centrifugation at 1100 x g for 10 min at 4 °C and subsequently stored at −80 °C. Samples from longissimus dorsi (LD) and superficial biceps femoris (BF) muscles were immediately collected into 2 ml reaction tubes. After opening the abdominal cavity, the entire liver was collected and a small aliquot taken into a 2 ml reaction tube. Subsequently, all tissue samples were snap-frozen in liquid nitrogen and stored at −80 °C pending analysis.