Fig. 3.

Specific pancortin isoforms reduce β-secretase- but not α-secretase-mediated cleavage of APP. HEK293 cells were transiently transfected with pancortin isoforms and individual domains or empty vector (ctrl). Twenty-four hours post-transfection, media were replaced, and after 24 additional hours media were collected and cells lysed. (A,D) Quantification of endogenous APPsα and APPsβ in the conditioned media (CM) via a multiplex ELISA, which allows for the simultaneous detection of each. (B) Quantification of Aβ40 in the CM by a separate ELISA. (C,E) Western blots of lysates (and conditioned media where noted by ‘CM’) showing expression of endogenous full-length APP, transfected pancortins and each of the secretases. Asterisk in C indicates a non-specific band. In A,B,D, each bar represents data from two to four independent experiments performed in triplicate for A and B, and a single representative experiment performed in triplicate for D. Error bars represent s.e.m. ***P<0.001.