Reducing Amyloid Plaque Burden via Ex Vivo Gene Delivery of an Aβ-Degrading Protease: A Novel Therapeutic Approach to Alzheimer Disease (original) (raw)
Figure 2
Clearance of the Aβ Peptide by NEP and sNEP
(A) CHO cells overexpressing APP were cocultured with CHO cells stably expressing either empty vector, NEP, or sNEP constructs. Equal numbers of cells were seeded, grown to confluence, and media were conditioned for 18 h. Conditioned media were analyzed for total Aβ levels by ELISA.
(B) Cellular lysates from the coculture in (A) were probed by Western blotting for APP (which presents in both mature and immature glycosylated forms), demonstrating equal amounts of APP expression. Each condition is shown in duplicate.
(C) Conditioned media from CHO cells stably transfected with APP were incubated in vitro with conditioned media from CHO cells expressing empty vector, NEP, or sNEP. The conditioned media were combined and incubated at 37 °C for 18 h. Aβ levels were determined by ELISA.
The immunoblot for APP (B) is representative of four experiments; ELISA values for Aβ represent the mean ± SEM of five experiments. For comparisons to the empty vector condition, ** p < 0.01 and *** p < 0.001.