Fig. 3.

Electron micrographs of liver from calorie-restricted WT (A–D) and Goat^−/− (F–I) mice at 9:30 AM and 5:30 PM on day 8. WT and Goat^−/− littermates (8 wk old) were subjected to 60% calorie restriction for 8 d. Mice were perfused at 9:30 AM or 5:30 PM on day 8 of calorie restriction, after which the liver was fixed and processed as described in Materials and Methods. Pictures from 20 different unit areas (36 × 26 μm per unit area) from two WT and two Goat^−/− livers at each time point were taken at 5,000 amplitude; representative pictures are shown. White arrows denote typical autolysosomes. (Scale bar: 5 μm; magnification, 5,000×.) (Insets) Enlarged images of each arrow-denoted autolysosome (magnified 5×). (E and J) The number of autolysosomes per unit area was determined by three independent examiners as described in Materials and Methods. Each value represents mean ± SEM of data from 20 images. Asterisks denote level of statistical significance (Student t test) between autolysosome numbers at 9:30 AM and 5:30 PM of WT (E) and Goat^−/− (J) mice: ***P < 0.001.