Fig.1 Western blot analysis of APPΔC31 (ref.3).
Human NT2 neurons (neurally differentiated human NT2 embryonic carcinoma cells) were infected with adenovirus vector expressing β-galactosidase (lane 1), wild-type APP (lane 2) or APPΔC31 (lane3). Cell lysates were prepared 48 h after infection, and proteins were analyzed by Western blotting using this antibody (SAC). Neurons overexpressing wild-type APP contained a 96 kD SAC-immunoreactive fragment which was also detected in APPΔC31-overexpressing neurons.
Fig.2 Immunocytochemical analysis of APPĒC31: Caspase-3 activation and generation of the caspase-cleaved fragment APPĒC31 within neurons induced by serum deprivation (ref.3).
Neurally differentiated NT2 cells were cultured for 96 h in the absence of fetal calf serum. Cells were triply labeled for the neuronal marker microtubule-associated protein 2 (MAP2), chromosomal DNA (Hoechst), and activated caspase-3 (ACP3; upper panel) or APPĒC31 (SAC; lower panel). MAP2-immunopositive neurons with apoptotic nuclei (arrows) are intensively immunostained with ACP3 and SAC.