FIGURE 7.
Reduced mineralization and altered gene expression by direct deletion of menin in osteoblasts. Primary calvarial Men1f/f osteoblasts were infected with adenovirus expressing GFP (control) or Cre recombinase. A, mineralization was assessed by Alizarin Red-S assay. i, staining at 21 days after culture in osteogenic medium. ii, quantitation. ***, p < 0.001; Adeno-Cre versus Adeno-GFP. B, RNA was isolated, semiquantitative RT-PCR and gel electrophoresis was performed, and densitometric analysis was made. Expression of osteoblast markers Runx2, Osx, and osteocalcin was reduced, the cyclin-dependent kinase inhibitor p27 was reduced, CDK4 was increased, osteocyte markers sclerostin and DMP-1 were increased, pro-apoptotic marker Bad was increased, and anti-apoptotic marker Bcl-2 was decreased. **, p < 0.01; ***, p < 0.001; Adeno-Cre versus Adeno-GFP.