Fig. 1.

Levels of blood glucose (A), plasma GH (B), and hepatic LC3-II (C–E) before and after feeding in calorie-restricted WT and Goat^−/− mice. Sixteen male WT and 16 Goat^−/− littermates (8 wk old) were subjected to 60% calorie restriction for 8–9 d as described in Materials and Methods. Four mice from each group were killed at the indicated time. (A and B) At 1 h before sacrifice, mice were injected i.p. with leupeptin (15 mg/kg). Immediately before sacrifice, blood was obtained from the retro-orbital sinus for measurement of blood glucose and plasma GH. Each value in A and B represents mean ± SEM of data from four mice. Asterisks denote the level of statistical significance (Student t test) between WT and Goat^−/− mice: **P < 0.01; ***P < 0.001. (C–E) Liver supernatants were processed as described in Materials and Methods, after which individual (C) or pooled (D) supernatants were subjected to immunoblotting with the indicated antibody. For immunoblotting of LC3 and GAPDH, 20 μg of liver supernatant was loaded in each lane; for p-STAT5 and t-STAT, 60 μg was loaded. Each pooled sample in D was derived from equal amounts of the four individual samples in C. (E) Relationship between the amounts of LC3II and p-STAT5. The relative amounts of LC3II (normalized to GAPDH) and p-STAT5 (normalized to total STAT5) were quantified by scanning the autoradiograms in D. The data were analyzed with ImageJ.