Fig. 2: TIMP2 is necessary for adult neurogenesis in the normal dentate gyrus.

A Schematic timeline of BrdU intraperitoneal injection protocol to label proliferating cells in the DG in isolated brain sections. B Schematic timeline of BrdU intraperitoneal injection protocol used for cell fate “survival” labeling in the DG of isolated brain sections. C Representative confocal microscopy images of BrdU⁺ cells in the DG of WT and TIMP2 KO mice (2–3 months of age, N = 11–12 mice per group; arrowheads indicate BrdU⁺ cells; scale bar, 50 μm) with corresponding (D) quantification of number of BrdU⁺ cells in the DG per unit area. E Representative confocal microscopy images of proliferating Ki67⁺ cells in the DG of WT and TIMP2 KO mice (2–3 months of age, N = 11–12 mice per group; arrowheads indicate Ki67⁺ cells; scale bar, 50 μm) with corresponding (F) quantification of the number of Ki67⁺ proliferating cells per DG in WT and TIMP2 KO mice. G Representative confocal microscopy images of Sox2⁺ cells in the DG of WT and TIMP2 KO mice (2–3 months of age, N = 11–12 mice per group; arrowheads indicate Sox2⁺ cells; scale bar, 50 μm) with corresponding (H) quantification of Sox2⁺ neural progenitor cells in the DG of WT and TIMP2 KO mice. I Representative confocal microscopy images of DCX⁺ cells in the DG of WT and TIMP2 KO mice (2–3 months of age, N = 11–12 mice per group; scale bar, 50 μm) with corresponding (J) quantification of DCX⁺ immature neuroblasts in DG of WT and TIMP2 KO mice. K Representative confocal microscopy images of BrdU⁺ NeuN⁺ cells in DG of WT and TIMP2 KO mice (2–3 months of age. N = 8–11 mice per group; scale bar, 100 μm and 50 μm (inset)) with corresponding (L) quantification of newborn neurons (BrdU⁺ NeuN⁺ cells) in the DG of WT and TIMP2 KO mice. Data are represented as mean ± SEM. Student’s t test for two-group comparisons (with Welch’s correction for (H) and (L)). *P < 0.05; **P < 0.01; ***P < 0.001. Data points represent individual mice. IHC immunohistochemistry, DG dentate gyrus, DCX doublecortin.