Murine Dietary Intervention and Tissue Sampling

Female HNKO and WT mice (10–16 weeks of age) were randomized into four groups based on lean mass. Mice were either fed an L-amino-acid defined, low MCD (A06071302; Research Diets) (39 (link)) or a matched, purified, L-amino acid defined low-fat PD (A06071314; Research Diets) for 3 weeks. Body composition was determined before and after the diet intervention using an NMR scanner (EchoMRI 4-1; EchoMRI). Before sacrifice, mice were fasted for 4 h from ZT = 3. Mice were sedated with pentobarbital as previously described (16 (link)) and killed by cardiac puncture with a heparin-coated needle. Blood was transferred to tubes with EDTA and was centrifuged at 6000g at 4 °C for 6 min. Plasma was transferred to a new tube and stored at −80 °C. Part of the hepatic median lobe was fixed in 4% paraformaldehyde (PFA) for histology. The remaining liver was snap frozen and stored at −80 °C. The frozen liver was crushed in liquid nitrogen before further analysis.