Blood samples were collected via the retro-orbital sinus using heparinized capillary tubes (2501; KILBLE CHASE, USA). Plasma insulin was measured using an insulin ELISA kit (#90080; Crystal Chem) according to the manufacturer’s instructions. Plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were analyzed at the Laboratory Animal Resource Center (KRIBB, Daejeon, Korea). Lipids were extracted from liver tissues using a chloroform/methanol mixture (2:1 v/v), as described previously19. Hepatic triglyceride (TG) was measured using a Triglyceride Determination Kit (TR100; Sigma–Aldrich).
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