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. 2006 Nov 27;27(4):1321–1333. doi: 10.1128/MCB.01280-06

FIG. 5.

FIG. 5.

Interaction and colocalization of endogenous ERα, Fe65, and APP in brains of APP and presenilin 1 double-transgenic (TG) and nontransgenic (non-TG) mice. (A) HeLa cells were transfected with 2 μg of pLENhERα and 4 μg of HA-Fe65, together with 4 μg of APP or APP mutant (APP*), and treated for 72 h. The interaction between full-length APP and ERα was determined by coimmunoprecipitation. (B to D) Coimmunoprecipitations were performed with homogenized brain tissues of female mice. The levels of endogenous proteins in mouse brain tissues or the precipitates were determined by immunoblotting. Extracts of HeLa cells transfected with control or ERα, Fe65, and APP expression vectors were included as controls. One mouse brain tissue was used per assay, and the data were reproduced twice. (E) Imaging analyses of female mouse brain sections through the area of the hippocampus. (E1) Coronal section through the hippocampus, with high-intensity Fe65 signal. (E2) Confocal images of tissue sections of a selected hippocampus area (see the white square in panel E1) after double staining for endogenous Fe65 (red) and ERα (green). (E3) Confocal images of tissue sections of the same hippocampus area after double staining for endogenous APP (red) and ERα (green).