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Fig. 3. Decreased bone formation in Smad4 mutant mice. (A,B) Von Kossa staining of the proximal tibia sections of Smad4 mutant (B) and control mice (A). (C,D) Representative images of the femora of 4-week-old male Smad4 mutant and control mice labeled with sequential doses of calcein. (E,F) Formation of mineralized ECM in neonatal calvarial osteoblast cultures from Smad4 mutant and control mice. Mineralized nodules were stained with Alizarin Red. (G-L) Quantitative histomorphometric measurements were performed on the secondary spongiosa at the distal femur of 8-week-old male mice. (G) BV/TV, bone volume/tissue volume; (H) TbN, trabecular number; (I) TbSp, trabecular separation; (J) ObS/BS, osteoblast surface/bone surface; (K) MAR, mineral apposition rate; (L) BFR/BS, bone formation rate/bone surface. All values are mean ± s.d. from six control (white bars) or Smad4 mutant mice (grey bars). ^*P<0.05; ^**P<0.01. Bar, 100 µm (A,B); 6.25 µm (C,D).

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