Figure 5:

Osteogenic cultures of Tg mice and littermate controls. (A) Calvarial osteoprogenitors isolated from wt and Tg mice were cultured in osteogenic medium and stained for ALP expression on day 7, 14 and 21, scale bar 50 μm. (B) ALP activity was measured using a colorimetric assay and expressed as absorbance normalized to the cell number determined by MTT assay. (C) Relative mRNA expression of Alpl, Runx2 and Osterix in osteogenic cultures on day 7 determined by real-time PCR, fold changes expressed relative to the expression in wt osteogenic cells. (D) Calvarial osteogenic cells from wt and Tg mice treated with 5 U and 10 U of EPO for 24 hours and ALP activity measured using colorimetric assay, absorbance was normalized to the cell number. (E) Calvarial osteoprogenitors from wt and Tg mice stained for mineralization using alizarin staining after 28 days of culture in osteogenic medium, scale bar 50 μm. (n=4/group, *p<0.05, **p<0.01).