Figure 2.

Tsc1 loss causes an accumulation of autolysosomes. Data shown are from rd1-mutant mice harboring the Tsc1^c/c allele at 2 months of age. (a) Immunofluorescence analyses on retinal flat mounts for proteins indicated (red signal). Cone layer was identified by PNA staining (green). (b and c) Immunofluorescence analyses on retinal flat mounts of retinae infected with the AAV9-mCherry-GFP-LC3 vector at birth. (b) Representative images at 2 months of age showing increased mCherry⁺/GFP^– punctae (arrows) in cones of Cre⁺ mice indicating an increase in number of autolysosomes. Arrowheads indicate GFP⁺/mCherry⁺ autophagosomes. (c) Bar graphs representing average number GFP⁺/mCherry⁺ punctae (autophagosomes) and GFP^-/mCherry⁺ punctae (autolysosomes) per cone cell. Data are representative of measurements in at least 60 cones over three different animals per genotype (****P<0.0001 by Student's t-test). (d) Immunofluorescence analyses (red signal) on retinal flat mounts for phosphorylation sites on indicated proteins. Cones were detected by SW OPSIN (green signal) staining. In all panels blue is nuclear DAPI. Scale bars: 20 μm