Fig. 2
Wnt1G177C/G177C mice display a bone-specific phenotype. a Representative images and quantification of alizarin red staining of ST2 cells transfected with empty vector (pLNCX) and expression plasmids for wild-type (Wnt1) or mutant WNT1(G177C) after 15 days of osteogenic differentiation. n = 3 samples per group. b Expression analysis by qRT–PCR of the indicated genes in ST2 cells transiently transfected with the same plasmids 5 days after transfection. n = 3 samples per group. c Representative Nissl staining of cortex, hippocampus, and cerebellum sections from 4-week-old Wnt1+/+ and Wnt1G177C/G177C mice. d Representative immunofluorescent images of the hippocampus in the same mice. Red: Anti-doublecortin (DCX); Blue: DAPI. e Quantification of the cortical Cux1-positive cell layer thickness and of the doublecortin-positive neuroblasts in the subgranular zone of the hippocampus. n = 3 animals per genotype. f Representative contact radiographs of spontaneous fractures in 24-week-old Wnt1G177C/G177C mice. g Number of spontaneous fractures observed per animal in female and male Wnt1+/+, Wnt1+/G177C, and Wnt1G177C/G177C mice at 12 and 24 weeks of age. Each symbol represents an individual mouse. h Histological images of a tibial fracture callus from a 24-week-old Wnt1G177C/G177C mouse. Top left: von Kossa/van Gieson stain; bottom left: Masson-Goldner stain, squares indicate areas magnified on the right side as indicated by the numbers. Top right: Masson-Goldner stain; bottom right: toluidine blue stain. i Representative images of tibial bone marrow from nonfractured 24-week-old wild-type or Wnt1G177C/G177C mice and quantification of the number of bone marrow adipocytes per marrow area (N.Ad/M.Ar). The red rectangle on the symbolic tibia in the top left corner indicates the region depicted in the representative images. Arrows and boxes indicate regions with high numbers of adipocytes. Data were analyzed by one-way ANOVA with Dunnett’s multiple comparison test (a, b) or Student’s t-test (e, i). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1. Error bars indicate standard deviation
