Fig. 4.

Ars2 is required for adult neurogenesis and select behavioral functions. (A) Immunostaining of adult dentate gyrus (DG). Bottom panels show magnification of boxed area, which indicates a SOX2⁺ cell with low Ars2. Dashed circles indicate co-localization of Ars2 (green), SOX2 (blue) and GFAP (red). (B) Adult DG shows co-localization of Ars2 (green) and DCX (red), but Ars2 is downregulated in some DCX⁺ cells (marked with asterisks). (C) Adult DG shows co-expression of Ars2 (green) and NeuN (red). (D) Scheme of tamoxifen administration in Nestin-CreERT2; Ai9; Ars2^fl/fl mice. (E-F′) Reduction of immature DG neurons co-expressing DCX (green) and tdTomato (red) in Nestin-CreERT2; Ars2^fl/fl; Ai9 mice compared with control littermates. E′ and F′ show magnifications of the boxed areas in E and F, respectively. (G) Quantification of E,F; n=3, two tailed t-test; data are mean±s.d. (H) Experimental scheme of proliferating NSC quantification assay used in J and K. (I,J) Quantitative evaluation of CldU⁺/SOX2⁺ NSCs and CldU⁺/IdU⁺ proliferating NSCs in the DG of control and induced cKO mice. *P<0.05. n=3, two-tailed t-test; data are mean±s.e.m. (K) Reduced immobility time of both male and female Ars2-CKO mice. C-TAM, control Ars2^fl/fl injected with TAM; C-W, control wild type; E-TAM, Cre⁺ Ars2^fl/fl injected with TAM. Repeated measures of ANOVA. Males: group effect F(2, 32)=4.449, P=0.0197; group×time interaction F(4, 64)=2.63, P=0.0424; multiple t-test with the Holm-Sidak method; C-W versus E-TAM *P=0.0081. N=6, 12, 13. Females: group effect F(2, 38)=4.08, P=0.0248; C-W versus E-TAM *P=0.0237 and 0.0306; C-TAM versus E-TAM ^#P=0.0179. N=8, 19, 14. Scale bars: 50 μm in A-C; 100 μm in E,F.