Fig. 4
From: Erythropoietin modulates bone marrow stromal cell differentiation
Osteogenic cultures of Tg6- and ΔEpoRE-mice. a Calvarial osteoprogenitors isolated from wt and Tg6-mice were cultured in osteogenic medium and stained for ALP expression on day 7 and mineralization deposits by alizarin red staining on day 21. b Relative expression of Alp, Runx2 and Osterix in osteogenic cultures on day 7 determined by real-time PCR. c Cell proliferation of calvarial osteoblasts was determined using MTT assay at 24, 48, and 72 h of culture in osteogenic medium and data expressed as absorbance values. d ALP activity was measured using a colorimetric assay and expressed as absorbance normalized to the cell number determined by MTT assay. e Calvarial osteoprogenitors isolated from wt and ΔEpoRE-mice cultured in osteogenic medium and stained for ALP (day 7) and alizarin red (day 21). f Relative expression of Alp, Runx2 and Osterix mRNA levels from primary calvarial osteogenic cells (day 7) determined by real-time PCR. g–h Calvarial osteogenic cells from wt and ΔEpoRE-mice treated with 5 U and 10 U of EPO and measured for their cell proliferation rate by MTT assay (g) and ALP activity of the cells were measured using colorimetric assays, absorbance was normalized to the cell number (h). (n = 4/group, **P < 0.01, ***P < 0.001)
