Figure 7. Decreasing the ubiquitination of APP alters its subcellular localization in primary neurons.

a, b) Mouse neurons were transfected with human APP_wt^GFP (in a) and APP_3R^GFP (in b) plasmids at DIV9. After 24h expression in the presence (right panels) or absence (left panels) of γ-secretase inhibitor compound XXI, neurons were fixed, stained for EEA1 (red channel) and imaged with confocal microscopy. Scale bar = 10 µm. Arrowheads indicate structures where APP^GFP and EEA1 colocalize

c) Bar diagram showing the amount of colocalization between GFP and EEA1 per 2500 µm² of image cell surface area following expression of APP_wt^GFP and APP_3R^GFP in cultured neurons, in the presence or absence of γ-secretase inhibitor XXI. Results are shown as means ± SEM (APP_wt^GFP, n=30 and APP_3R^GFP, n=20). ***, denotes P values < 0.001 from a Student's t-test.