Figure 3.

Identification of autophagosomes and reduction of mTOR activity in the cerebellum of food-restricted mice. (A) Low magnification flattened images of GFP-LC3 fluorescence were re-assembled to generate a complete sagittal cross-section of cerebellum from normal and food-restricted mice (left column). Higher magnification images of the white boxed regions show GFP-LC3 fluorescence in purkinje cells along the boundary between the molecular and granular layers (second column). Additionally magnified representative images of the (boxed) Purkinje cell bodies reveal finer detail and localization of the GFP-LC3 signal (far right column). A merged fluorescent image for each mouse is shown in the third column; GFP-LC3 (green), nuclei (blue), pan-neuronal marker (orange), GFAP/astrocyte marker (red). (B) Cerebellar sections were stained with an antibody specific for phospho-S6RP, a protein whose abundance varies directly with mTOR activity, and inversely with autophagy (see text).