Supplementary Figure 1: Construction and characterization of ALK5cKO transgenic mice.
From: ALK5-dependent TGF-β signaling is a major determinant of late-stage adult neurogenesis
(a) Diagram of gene constructs used to create ALK5cKO mice. (b,c) Immunoblot of hippocampal lysates from 3-month-old ALK5loxP/loxP and ALK5cKOmice probed with antibodies against ALK5, NSE, and actin in (b) and quantification of ALK5 signal intensity normalized against NSE in (c), n = 2 mice per group, P = 0.0276. Full-length blots are presented in Supplementary Figure 10. The experiments were independently performed twice. (d,e) Representative tiled confocal images (d) and quantification (e) of the dentate gyrus from 3-month-old ALK5loxP/loxP (n = 8) and ALK5cKO(n = 7) mice immunostained for p-Smad2. P-Smad2 signals were normalized to background in (e). Three sections per mouse, P = 0.0433. (f) Representative orthogonal projections from confocal Z-stack images of the dentate gyrus from 3-month-old mT/mG mice before and after crossing with CamK2a-cre mice immunostained for eGFP in combination with Sox2 plus GFAP, MCM2, Tbr2, DCX plus PCNA, and NeuN. DAPI stains nuclei. Scale bar is 100 μm in (d) and 10 μm in (f). (g) Quantification of eGFP positive cells in different cell types detected in (f) from the dentate gyrus. (h) Quantification of eGFP positive and negative cells in individual cell types detected in (f) from the dentate gyrus. n = 4 mice, 3 sections per mouse in (g,h). Data are presented as mean ± s.d. in (c), mean ± s.e.m. in (e,g,h). *P < 0.05, ****P < 0.0001 Student's t-test in (c,e); one-way ANOVA, Dunnett's post-hoc test in (g).
