Additional Supporting information may be found in the online version of this article.

JBMR_272_sm_SuppFig1.tif7648KSupplemental Figure 1. Femurs harvested from EP1−/− and wild-type control mice (N=5 mice/group) at days 7, 14 and 21 post-fracture were scanned at 10.5 micron isotropic resolution using a Scanco VivaCT 40 (Scanco Medical AG) (A). Reconstruction of µ-CT data were collected from the fracture callus region of the femur as previously described (24). The fracture callus bone volume and mineral density were determined (B). Data are presented as mean ± SD. Statistical comparisons were performed using 2-way ANOVA with Bonferroni post-hoc multiple comparisons. “a” indicates significant differences compared to wild-type (p < 0.05).
JBMR_272_sm_SuppFig2.tif761KSupplemental Figure 2. Femurs were dissected at day 28 post-fracture from EP1−/− and wild-type mice (N=5 mice/group). Torsion testing was performed on a Torsional Test Bench (Endura TEC) according to established protocols (45). Briefly, the femurs were dissected free of muscle and soft tissue, and the pins were carefully removed without disrupting the callus. The ends of the femurs were cemented into 6.35 mm2 aluminum tube holders using PMMA in a custom jig to ensure axial alignment and to maintain a gage length of ∼6.5 mm, allowing at least 3 mm to be potted at each end. Specimens were bathed in PBS at room temperature for at least 2 h after potting to allow for rehydration of the tissue and hardening of the PMMA. Specimens were then mounted on an EnduraTec TestBenchTM system (200 torque cell; Bose Corporation, Minnetonka, MN) and tested in torsion at a rate of 1°/s until failure. The torque data were plotted against the rotational deformation (normalized by the gage length and expressed as rad/mm) to determine the (A) ultimate torque, (B) torsional rigidity (slope of the linear region of the torque-normalized rotation curve), (C) ultimate Rotation, and (D) energy to failure (area under the torque-normalized rotation curve). Statistical comparisons were performed using unpaired Student's t-test with Welch's corrections (does not assume equality of variances) after verifying normality of the data using the Kolmogorov-Sminrof test. “a” indicates significant differences compared to wild-type (p < 0.05).

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