Fig. 5
The subcellular localization of APP-CTFs is different in PS+/+ and PS1/2−/− MEF cells. a MEF PS+/+ cells transfected with C99 or C99G29L/G33L and treated or not with D6, or MEF PS1/2−/− cells were stained with α-APPct. Whereas vehicle-treated C99-transfected MEF PS+/+ cells presented Golgi-like staining, D6-treated cells, aswell as C99G29L/G33L expressing and PS1/2−/− cells, displayed mainly punctuate staining. b MEF PS1/2−/− cells were co-transfected with C99 and the intracellular markers Rab5-GFP, Rab7-GFP or Lamp1-GFP (green) and immunostained with α-APPct (red). The right panels show 2-fold high magnification of merged images. c Bars correspond to the quantification of cells having either Golgi/ER or endosome/lysosome associated APP-CTF immunostaining. A total of 400 cells for each condition from 3 independent experiments were counted. Data represent means ± SEM, ***p < 0.001 as analyzed by 2-way ANOVA followed by Dunetts posthoc analysis. d Immunoblot analysis of APP-CTFs in MEF PS+/+ and PS1/2−/− cells. Proteins were stained by photoactivation using the Bio-Rad prestain method (protein stain) and immunoblots used for the detection of APP-CTFs. Arrows point to HMW immunoreactivities