Figure 6.

Accumulation of membrane-tethered APP intracellular domain-induced neurite outgrowth requires adenylate cyclase activation. N2a cells and cortical neurons coexpressing YFP and EV, mCtl, or mAICD were stimulated with FSK (1 μm; 24 h) and/or treated with adenylate cyclase inhibitor MDL-12,330A (10 nm; 24 h), and neurite extensions were quantified as described in Materials and Methods. Representative images of N2a cells (a) or inverted images of cortical neurons (c) are shown. b, d, Three-dimensional deconvolved images were quantified, and morphological changes following FSK stimulation and/or application of adenylate cyclase inhibitor MDL-12,330A (10 nm; 24 h) are plotted as relative difference of total neurite area compared with untreated EV or mCtl controls. Statistical analysis was performed using ANOVA Kruskal–Wallis test followed by Dunn's post hoc multiple-comparison analysis. *p < 0.05, **p < 0.001, compared with untreated EV- or mCtl-expressing cells; ^##p < 0.001, compared with untreated cells within the same transfected conditions. The total number of quantified cells is shown in parentheses. Error bars indicate SEM.