(A) Increased cell proliferation in FoxO3−/− neurospheres. Cells dissociated from secondary neurospheres from 3 month-old FoxO3+/+ or FoxO3−/− mice were plated on poly-D-lysine and incubated for 1 hour with BrdU. Cells were immunostained with antibodies to BrdU. BrdU-positive nuclei were counted. Values represent mean ± SEM from 2 independent experiments with 5 littermates for each genotype. Student's t-test, ***: p<0.001.
(B) Apoptosis in FoxO3−/− NSC compared to FoxO3+/+ NSC. Freshly isolated NSC or cells dissociated from secondary neurospheres isolated from 3 month-old FoxO3+/+ or FoxO3−/− mice were stained with antibodies to cleaved caspase 3. Cleaved caspase 3-positive cells were counted. Values represent mean ± SEM from 2 independent experiments with 5 littermates for each genotype. Student's t-test, **: p<0.01.