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

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jbmr2059-0001-sm-SuppFig-S1.eps1352K

Fig. S1. Longitudinal analysis of body weight and tibial length.

Body weight gain (A and B) and total tibial bone length (C and D) were measured at monthly or bi-monthly intervals in wild-type (WT, circles), Sost−/− (squares), Lrp5−/− (triangles) and Sost−/−;Lrp5−/− (inverted triangles) female (A and B) and male (C and D) mice. Data represent means ± SEM. Group size: n = 6–18 (females), n = 3–15 (males). Statistical significances determined by one-way ANOVA are shown for 6-month-old-mice and are designated as follows: +, p < 0.05 Sost−/−;Lrp5−/− versus Sost−/− mice.

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Fig. S2. Lrp5/6 expression in femoral cortical bone.

Lrp5 (A) and Lrp6 (B) gene expression normalized to 18S expression was measured by qPCR in femoral cortical bone of 6-month-old mice wild-type (WT, white bars), Sost−/− (grey bars), Lrp5−/−(hatched bars) and Sost−/−;Lrp5−/− (dark bars) mice and calculated relative to expression levels in wild-type (WT) mice. Data represent means plus SEM. Group size: n = 5–10 (females), n = 3–8 (males). Statistical significance determined by one-way ANOVA is designated as follows: *, p < 0.05; **, p < 0.01 versus wild-type (WT); +, p < 0.05; ++, p < 0.01 versus Sost−/−; x, p < 0.05; xx, p < 0.01 versus Lrp5−/− mice.

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Fig. S3. Serum osteocalcin in adult mice treated with anti-Lrp6 antibodies.

Serum osteocalcin as a marker for osteoblastic bone formation was determined in 16-week-old wild-type (WT, white bars) and Sost−/− (dark bars) female mice after one-month treatment with vehicle, anti-Lrp6 antibody A, the biparatopic anti-Lrp6 antibody A/B or anti-Lrp6 antibody B, respectively (Veh, A, A/B, B). Data represent means plus SEM. Group size: n = 7–10 (wild-type), n = 4–10 (Sost−/−). Statistical significance determined by two-way ANOVA is designated as follows: +, p < 0.05; ++, p < 0.01 versus antibody A-treated mice of the same genotype; x, p < 0.05; xx, p < 0.01 versus antibody A/B-treated mice of the same genotype; †, p < 0.05; ††, p < 0.01 versus mice of different genotype with the same treatment.

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Fig. S4. Serum serotonin in 3-month-old mice.

Serum serotonin concentration was determined in wild-type (WT, white bars), Sost−/− (grey bars), Lrp5−/−(hatched bars) and Sost−/−;Lrp5−/− (dark bars) mice. Data represent means plus SEM. Group size: n = 5–10 (females), n = 4–9 (males). No statistical significant differences were found as determined by one-way ANOVA.

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