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. 2022 Feb 4;11:e60183. doi: 10.7554/eLife.60183

Figure 6. Bone formation is not increased by Dll4(E12) in ovariectomized female mice.

(A, B) Representative 3D micro-computed tomography (μ-CT) images of trabecular bone in the distal tibial metaphysis (A) and cortical bone in the mid-tibial diaphysis (B) of sham or ovariectomized mice treated with vehicle or pLIVE-Dll4(E12). (C) Bone parameters measured by μ-CT analysis of trabecular bone volume/total volume, trabecular connectivity density, trabecular thickness, trabecular separation, bone surface, and trabecular number in the distal tibial metaphysis. Data represent mean ± SD. (n = 5 mice) (p-values determined by two-way ANOVA with Tukey’s multiple comparisons test). (D) Quantitation of μ-CT analysis of cortical bone consistency and cortical thickness in the mid-tibial diaphysis. Data represent mean ± SD. (n = 5 mice) (p-values determined by two-way ANOVA with Tukey’s multiple comparisons test). (E, F) Tile scan confocal images showing Collagen I alpha one chain (Col1a1) staining (green; E) and PDGFRβ staining (green; F) in femoral sections from ovariectomized female mice treated with vehicle or pLIVE-Dll4(E12). Nuclei, DAPI (blue). Graphs show relative ratio (percentage) of Col1a area/DAPI area and PDGFRβ area/DAPI area. Data represent mean ± SD. (n = 5 mice) (p-values determined by two-tailed unpaired t-test (E) with Welch’s correction (F)). (G) Representative μ-CT images of trabecular bone in the distal tibial metaphysis of male, female sham, and ovariectomized mice. Graphs show quantitation of the trabecular bone volume/total volume, trabecular number, and bone surface. Data represent mean ± SD. (n = 5 mice), (p-values determined by one-way ANOVA followed by Sidak’s multiple comparisons test).

Figure 6—source data 1. Source data for Figure 6C–G.

Figure 6.

Figure 6—figure supplement 1. Reduced Dll4(E12) immunoreactivity and trabecular bone in female mice compared to male mice.

Figure 6—figure supplement 1.

(A, B) Representative overview confocal images of Collagen I alpha one chain (Col1a1) staining (green) in the femurs of wild-type male and female mice treated with vehicle (A) or pLIVE-Dll4(E12) (B). Nuclei, DAPI (blue). Data represent mean ± SD. (n = 5–7 mice) (p-values determined by two-tailed unpaired t-test (control) or unpaired t-test). (C, D) Representative overview confocal images of Dll4 staining (green) in the femurs of wild-type male and female mice treated with vehicle (C) or pLIVE-Dll4(E12) (D). Nuclei, DAPI (blue). Diagrams represent percentage of Dll4 area/DAPI area. Data represent mean ± SD. (n = 5–7 mice) (p-values determined by two-tailed unpaired t-test (control) or unpaired t-test with Welch’s correction (Dll4(E12))).
Figure 6—figure supplement 1—source data 1. Source data for Figure 6—figure supplement 1A-D.