(A) FoxO3−/− NSC from adult mice display a defect in primary neurosphere formation. NSC isolated from 3 month-old FoxO3+/+ or FoxO3−/− littermates were seeded at the indicated cell densities. The number of neurospheres formed after one week was counted. Values represent mean ± SEM from 7 independent experiments performed with 5 littermates for each genotype. Two-way ANOVA (p<0.0001 for the genotype variable), Bonferroni post-tests, ***: p<0.001.
(B) FoxO3−/− NSC from adult mice, but not from neonates, display a defect in primary neurosphere formation. Frequency of primary neurospheres formed from NSC at low-density (8,000 cells/ml) from mice at different ages (1 day-old, 1d; 3 month-old (3m); 1 year-old (1y)). Values represent mean ± SEM from 2 independent experiments with 2 littermates (1d), 7 independent experiments with 5 littermates (3m), and 3 independent experiments with 3-5 littermates (1y) for each genotype. Two-way ANOVA with Bonferroni post-tests, *: p<0.05; ***: p<0.001.
(C) FoxO3−/− NSC from adult mice, but not from neonates, display a defect in secondary neurosphere formation. Dissociated cells from primary neurospheres from FoxO3+/+ or FoxO3−/− littermates were seeded at 4,000 cells/ml. The number of secondary neurospheres formed after one week was counted. Values represent mean ± SEM from 2 independent experiments with 2 littermates (1d), 4 independent experiments with 5 littermates (3m), and 4 independent experiments with 3 to 5 littermates for each genotype (1y) for each genotype. Two-way ANOVA with Bonferroni post-tests, *: p<0.05.