Figure 4
Sod2 loss in osteocytes decreases the bone-forming activity associated with the upregulation of sclerostin.
(a) Fluorescent micrographs of representative calcein-double labeling of Sod2ot/ot and wild-type male femurs at 5 months of age. The mineralized surface per bone surface (MS/BS), mineral apposition rate (MAR), bone formation rate (BFR/BS), osteoblast surface per bone surface (Ob.S/BS) and number of osteoblast per bone surface (N.Ob/BS) (n = 4–5 each group). (b) Decalcified femoral sections were prepared from Sod2ot/ot and wild-type male femurs at 5 months of age and subjected to sclerostin immunohistochemical staining. (c) A gene expression analysis of osteoblast- and osteocyte-related genes in the flushed tibia at 5 months of age. The data were normalized to the Gapdh expression (n = 5 each group). (d) A Western blot analysis of the sclerostin levels in the flushed tibia at 5 months of age using the same gel as in Figure 3d. (e) Mineralized nodule formation in the culture of primary bone-forming cells derived from neonatal calvaria (upper panel) and bone marrow (lower panel) on culture day 28. *p < 0.05. The error bars indicate the SD. The scale bar indicates 50 μm.
