FIGURE 1.

Analysis of APP levels and proliferation of NSPCs derived from Tg2576 mice and background strain C57Bl/6xSJL mice. A, Western blot analysis of the level of APP in the conditioned medium of neurosphere cultures. Blots were stained with mAb 22C11, which recognizes both human and mouse APP, and mAb 6E10, which is specific for human-sequence APP. B, analysis of the proliferation of NSPCs. Neurospheres were dissociated, and the isolated cells were seeded on poly-l-lysine-coated 96-well plates at a density of 2000 cells/well. After 2, 4, or 6 days in culture, the relative number of cells was estimated using the alamarBlue assay. Fluorescence intensity was taken as an index of cell number. Values are means ± S.E. (n = 3 wells). C, phase-contrast microscopy of neurospheres after 7 days in culture. Scale bar = 200 μm. D, quantitation of the area of neurospheres from the experiment shown in C. Values are means ± S.E. (n = 200 neurospheres). E, EdU incorporation assay of cell proliferation. NSPCs were cultured in proliferation medium for 4 days prior to incubation for 8 h with EdU (30). The graph shows the mean values (± S.E.) of the percentage of cells incorporating EdU (n = 30 incubations). * = significantly different from corresponding values for the background strain C57BL/6xSJL cells (p < 0.05, experiment in panel B, one-way ANOVA with post hoc Tukey's test; experiment in panel D, Student's t test).