C57bl 6 000664

C57BL/6 (000664), Rictor^flox/flox (020649), Raptor^flox/flox (013188) and Tsc1^flox/flox (005680) mice were from Jackson Laboratory. Studies were performed in 2–10-month-old male and female mice fed regular chow (5058; Lab Diet) and maintained in barrier facility at 22–23 °C under 40–60% humidity and a 12 h:12 h light/dark cycle. Liver-specific Rictor^KO, Raptor^KO or Tsc1^KO mice were generated via retro-orbital injections of 2 × 10¹¹ genome copies per mouse of AAV8-TBG-iCre adenovirus (Vector Biolabs, VB1724) and mice were humanely killed after 8 weeks^{64 (link)}. AAV8-TBG-eGFP (Vector Biolabs, VB1743)-injected mice were controls. Mice were fasted with free access to water and were compared with ad libitum mice. Fasted mice were treated with: oral gavage of (1) corn oil (400 μl; Sigma-Aldrich, 8267), (2) BODIPY FL C₁₆ (10 mg kg⁻¹; Invitrogen, D3821) or (3) refed high-fat diet (60% kcal in fat; Research Diets, D12492) for 30 min. Mice were mock-gavaged for 5 days before experiment, and control animals were gavaged with vehicle (10% dimethyl sulfoxide–saline solution) to match for volume and distension. STZ (40 mg kg⁻¹; Sigma-Aldrich, S0130) was injected intraperitoneally once a day for 5 consecutive days. The protocol was repeated after 8 weeks, and tissues were collected 2 weeks after the last injection.