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N2a mouse neuroblastoma cells stably transfected with the Swedish mutant form of APP (SweAPP N2a cells; APP695, 595–596 KM/NL) (gift from G. Thinakaran and S. Sisodia, University of Chicago, Chicago, Illinois, United States) were maintained in DMEM, 10% FBS, and 200 μg/ml G418 in the presence of penicillin and streptomycin [32].
Sisodia and colleagues [39] demonstrated that arrest of APP endocytosis from the plasma membrane by deletion of its NPXY clathrin-coated vesicle targeting sequence [40,41] can dramatically stimulate sAPP α shedding, presumably by extending the half-life of co-localized α-secretase and APP on the plasma membrane.