FIGURE 6.

Lumican deletion reduces calvaria bone formation. The calvaria bones dissected from timed-pregnant mice at E21.0 were infected with lumican shRNA CM in BGjb medium (containing 100 U/mL penicillin) as well as 100 μg/mL streptomycin and 0.1% BSA (1:1 ratio) for 3 days and additionally cultured in a medium containing 50 μg/ml insulin for 10 days. (A) Hematoxylin and eosin staining was performed on the specimens, and calvaria bone widths were quantitated by analyzing the average of five spots. (B) Toluidine blue O (blue) staining was performed on the specimens, and the number of osteoblasts (red arrows) was counted. Scale bars, 20 μm (A, B). Data are presented as mean ± SEM. *P < 0.05 vs. control.