Figure 8.

Cystatin B deletion in TgCRND8 promoting clearance of Aβ, ubiquitinated proteins and LC3-II in isolated autophagic-lysosomal compartments. Subcellular fractions containing the autophagic-lysosomal compartments AV10, AV20 and lysosomes were isolated from brains of 6-month-old TgCRND8 and CBKO/TgCRND8 (n = 8 per genotype). Equal amounts of proteins from each fraction were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis and processed for western blotting with an antibody directed against Aβ (A), ubiquitin (B) or LC3 (C), and representative blots are shown. (D) AV20 and Lyso fractions from all four genotypes were loaded onto one gel and processed with an anti-cathepsin D (RU2) or anti-LC3 antibody. Corresponding ponceau S-stained blots are shown below the antibody-probed blots as loading controls (A–D). AV10 = early autophagic vacuole/autophagosome; AV20 = late autophagic vacuole/autolysosome; CRND8 = TgCRND8; Cyto = cytosol; ER = endoplasmic reticulum-enriched; Homo = homogenate; Lyso = lysosome; WB = western blotting; WT = wild-type.