Fig 1.

CHIP prevents β-cleavage of APP through BACE1 destabilization. (A–B) Destabilization of ectopic BACE1 by CHIP (0.5, 1, 2, and 3) μg in a dose-dependent manner. (C) CHIP destabilizes endogenous BACE1 level in SH-SY5Y neuroblastoma cells. (D) Silencing of endogenous CHIP stabilizes BACE1 protein level. (E) and (F) Functional domain of CHIP responsible for BACE1 destabilization. (G–H) CHIP destabilizes BACE1 protein level and decreases APP processing at β-cleavage site. (G) HEK 293 cell stably expressing APP were co-transfected with Flag-BACE1 constructs along with increasing amounts of myc-CHIP constructs. (H) Rat primary cortical neurons were transfected with increasing amount of myc-CHIP. After 30 h of transfection, C-terminal β-site cleavage product of APP in whole-cell lysate was determined by Western blotting using anti-APP antibodies (C-terminal). All the data were expressed as mean ±SE from three independent experiments. Statistical analysis were performed by one-group t-test for the significance at the * = P < 0.05, ** = P < 0.01 and *** =P < 0.001.