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. Author manuscript; available in PMC: 2010 Nov 6.
Published in final edited form as: Cell Stem Cell. 2009 Nov 6;5(5):527–539. doi: 10.1016/j.stem.2009.09.014

Figure 5. Consequences of FoxO3 loss in the nervous system on brain weight and the NSC pool.

Figure 5

(A) Expression of FoxO3 in the tissues of FoxO3lox/lox;Nestin-Cre mice. FoxO3 expression in different tissues of 2 month-old FoxO3lox/lox;Nestin-Cre mice was determined by Western blot with antibodies to FoxO3 (‘NFL’) and antibodies to β-actin and karyopherin β. Note that FoxO3 was partially deleted in the pancreas.

(B) Brain weights of FoxO3lox/lox;Nestin-Cre and FoxO3lox/lox mice. Brain weights were measured after perfusion in 3 to 4 month-old mice (3-4m) and 1.6 year-old mice (1.6y). Values represent mean ± SEM from 9-10 males and 9-10 females (3-4m), and 1-3 males and 5-8 females (1.6y). One male and 2 females (3-4m) and 3 females (1.6y) of the FoxO3lox/+ genotype were included in the control group (FoxO3lox/lox).

(C) Quantification of label-retaining NSC in FoxO3lox/lox;Nestin-Cre and FoxO3lox/lox animals in vivo. Number of BrdU-positive cells in the SGZ (left panel) and the SVZ (right panel) in 3 month-old FoxO3lox/lox;Nestin-Cre and FoxO3lox/lox littermates injected daily with BrdU for 7 days and sacrificed one month after the last BrdU injection. The number of BrdU-positive cells in the SGZ was normalized to the volume of the granular cell layer (GCL). Values represent mean ± SEM (left panel) and mean ± SD (right panel) from 8 FoxO3lox/lox;Nestin-Cre mice and 11 FoxO3lox/lox control littermates. Two FoxO3lox/+ littermates were included in the control group (FoxO3lox/lox). Mann-Whitney test, p=0.27 (left panel) and p=0.14 (right panel).

(D) Ablation of the FoxO3 protein in NSC from FoxO3lox/lox;Nestin-Cre mice. FoxO3 expression in NSC isolated from 3 month-old FoxO3lox/lox mice (−) or FoxO3lox/lox;Nestin-Cre mice (+) seven days after isolation (NS1) or at 3 consecutive passages (NS2 to 4) was determined by Western blot with antibodies to FoxO3 (‘NFL’) and antibodies to β-actin.

(E) Ablation of FoxO3 in the brain impairs primary neurosphere formation. NSC isolated from 9 month-old FoxO3lox/lox mice (control) or FoxO3lox/lox;Nestin-Cre mice were seeded at low density. The number of neurospheres formed after one week was counted. Values represent mean ± SEM from triplicates from 4 independent experiments conducted with 3 littermates for each genotype. Twoway ANOVA, p<0.01 for the genotype variable, Bonferroni post-tests, **: p<0.01.