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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: J Bone Miner Res. 2013 Sep;28(9):1925–1935. doi: 10.1002/jbmr.1938

Figure 7. Decreased bone volume and osteoblast function in old mice is rescued in Wwp1−/−mice.

Figure 7

(A–C) BMSCs from young (1-month-old) and old (12-month-old) WT mice were used. (A) The area of alizarin-red-stained calcified nodules. (B) The % of CXCL12-mediated migration. Values are the means ± SD of 3 wells/group. *, p<0.05, **, p<0.01 vs data from young mice. (C) Expression of Runx2, JunB and CXCR4 proteins was determined by Western blot analysis and the fold changes were calculated as Fig. 1D. (D–F) BMSCs from old Wwp1−/− mice and WT littermates were used. (D) The area of alizarin-red-stained calcified nodules. (E) The % of CXCL12-mediated migration. Values are the means ± SD of 3 wells/group. *, p<0.05, **, p<0.01 vs data from WT littermates. (F) Expression of Runx2, JunB and CXCR4 proteins was determined by Western blot analysis and the fold changes were calculated as Fig. 1D. (G) BMSCs derived from Wwp1−/− mice and WT littermates were treated with MG 132 (10 μM) for 4 hours and then lysed for ubiquitination assays to detect endogenous ubiquitinated JunB and Runx2 proteins. (H) BMSCs from Wwp1−/− mice and WT littermates were transfected with JunB siRNA or control siRNA. Protein expressions of Runx2 and JunB were determined by Western blot analysis.