Figure 3. Bone becomes more mineralized with age, and with increasing distance from the growth plate in Dmp1^Cre mice; this is delayed in Dmp1^Cre: Socs3^f/f mice.

(A) The metaphyseal region was analysed in consecutive 9 micron slices. (B–E) Bone volume at low-, mid-, and high-density mineral levels in each slice from the distal to proximal end of the metaphyseal region at 12 weeks (B,D) and 15 weeks (C,E) of age in Dmp1^Cre (B,C) and Dmp1^Cre:Socs3^f/f mice (D,E). Values are mean+ SEM, n = 9–11 mice per group; arrows denote where data becomes significantly different to the furthest slice from the arrow; colours denote the density level of bone that has changed. Pseudo-colourised images based on the Otsu thresholds show a representative sample for the top and bottom slice for each graph. Differences between genotypes are shown in Figure 3—figure supplement 2. (F,G) Ploton silver stain for osteocyte canaliculi and cement lines at the base of the metaphyseal region cut through the femur as indicated in the inset images of C and E for Dmp1^Cre (F) and Dmp1^Cre:Socs3^f/f mice (G). Scale bar = 20 micron.

Figure 3—figure supplement 1. Total bone area of the metaphyseal slices in Dmp1^Cre and Dmp1^Cre:Socs3^f/f mice at 12 and 15 weeks of age (B,D,F).

Data are mean ± SEM; n = 9–11 mice per group.

Figure 3—figure supplement 2. Comparison of high density, (A,B), mid-density (C,D) and low density (E,F) bone between Dmp1^Cre and Dmp1^Cre:Socs3^f/f mice at 12 (A,C,E) and 15 weeks of age (B,D,F).

Data are mean with SEM shown by dashed lines, taken slice by slice through the metaphysis (see Figure 3A for schematic). Gray region indicates significant difference between genotypes by repeated measures two-way ANOVA with Šidák post-hoc test.

Figure 3—figure supplement 3. Direct comparison of archived scans from Dmp1^Cre and Dmp1^Cre:Socs3^f/f mice at 26 weeks of age.

(A) Bone volume, as a percentage of total metaphyseal volume, segregated by low, mid and high density volumes; changes in low-, mid- and high-density bone are indicated by coloured asterisks; error bars shown are SEM for the low-, mid- and high-density bone volumes. **, p<0.01; ***, p<0.001 for comparisons shown, determined by repeated measures two-way ANOVA with Šidák post-hoc test; n = 9 per group. (B–D) Comparison of high density (B), mid-density (C) and low-density (D) bone between female Dmp1^Cre and Dmp1^Cre:Socs3^f/f mice at 26 weeks of age. Data are mean with SEM shown by dashed lines, taken slice by slice through the metaphysis (see Figure 3A for schematic); n = 9 mice per group. Gray region indicates significant difference between genotypes by repeated measures two-way ANOVA with Šidák post-hoc test.( E,F) Pseudo-colourised images showing a representative sample for the distal and proximal slices for each female genotype.