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

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Supplemental Fig. 1. Longitudinal analysis of cortical parameters. (A) Histomorphometric analysis of the structural properties of the femurs showed significantly decreased cortical bone area at all ages and decreased marrow area at 4- 12 months in Gja1Jrt/+ versus WT mice. Solid and dashed lines indicate significant differences over time in WT and Gja1Jrt/+ mice, respectively; n ≥ 6. (B) There was no difference in the percentage of lacunae that were empty in the cortical bone of Gja1Jrt/+ versus WT mice at 2 and 8 months of age; n ≥ 4. *p < 0.05, **p < 0.01 and ***p < 0.001.

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Supplemental Fig. 2. Analysis of osteoblast and osteocyte-specific genes in cortical bone extracts. (A) Expression of early-, mid- and late-osteoblast and osteocyte differentiation markers were unaffected in RNA isolated from cortical bone of 2, 4, 8 and 12 month old mice, with the exception of decreased Sost expression at 2 months in Gja1Jrt/+ versus WT mice; n ≥ 3 and each sample is comprised of two or more independent biological samples. Samples were run in triplicate. *p < 0.05, **p < 0.01 and ***p < 0.001.

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Supplemental Fig. 3. Expression of Rankl-Opg and osteoclast-specific gene expression in Gja1Jrt/+ versus WT mice. (A) The Rankl/Opg expression ratio in RNA extracted from cortical bone at all ages was unaffected in Gja1Jrt/+ versus WT; n ≥ 3 and each sample is comprised of two or more independent biological samples. (B) Rankl/Opg expression ratio was also unaffected in trabecular bone at 2 and 4 months of age; n ≥ 4. (C) Expression of osteoclast differentiation and fusion markers was unaffected in RNA from trabecular bone of 2 and 4 month-old Gja1Jrt/+ versus WT mice; n ≥ 3. Samples were run in triplicate. *p < 0.05, **p < 0.01 and ***p < 0.001.

jbmr1965-sm-0004-SuppTab-S1.doc46K

Supplemental Table 1. Quantitative RT-PCR primer sequences used in this study.

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Supplemental Table 2. List of antibodies used in this study.

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