Fig. 7

Elevated number of proliferating NSCs in the SGZ of Tfeb-cKO mice. a aNSCs in Tfeb-cKO mice and control mice were labeled with BrdU for 2 h before fixation on day 6 or at 4 weeks (4 W) after tamoxifen administration. b Representative images of the DG in wild type (WT) and cKO mice (HOMO) at 4 weeks, immunostained with GFAP (blue), BrdU (green), tdTomat (red), and Sox2 (gray). c, d Dot plots showing the number of BrdU-positive aNSCs in the DG of wild type (WT), heterozygote (HET), and homozygote Tfeb-cKO (HOMO) mice. Cells were counted in each image from 50-µm-thick FF-IHC sections throughout the DG. Four and six slices from each brain were counted on day 6 (c) and at 4 weeks (d), respectively (n = 4 [WT], 3 [HET], and 3 [HOMO] for day 6; n = 3 [WT], 2 [HET], and 3 [HOMO] for 4 weeks). e Tfeb-cKO mice (HOMO) and control mice (HET) were analyzed 12 weeks after tamoxifen administration. f Representative images of the DG from the control (HET) and Tfeb-cKO mice (HOMO), immunostained for GFAP (blue), Ki-67 (green), and Sox2 (gray) and processed using the Imaris software (right panels). Processed images were used to count NSCs (magenta spots) and aNSCs (green spots) using the Imaris software. g Enlarged view of white squares in f. h, i Dot plots showing the numbers of total NSCs and aNSCs in slices. Six slices from each brain were counted (n = 4 [HET] and 3 [HOMO]). j, k Proportions of aNSCs to NSCs (j) and qNSCs to NSCs (k), calculated from (h) and (i). l Representative images of the DG, immunostained as indicated (left panels) and processed (right panels) to count DCX-positive cells (green spots) and NSCs (magenta spots). m Ratio of DCX-positive cells to NSCs. Six slices from each brain were counted (n = 2 [HET] and 3 [HOMO]). n, o Average number of cells double-positive for tdTomato and DCX (n) or tdTomato and cleaved-caspase-3 (o) in the DG in slices. Data represent means ± s.e.m. (*P < 0.05, **P < 0.01, ****P < 0.001, *****P < 0.0001; Student’s t-test). Scale bars, 200 µm.