Figure 4

Expression of key transcription factors during differentiation of equine bone marrow stem cells into osteoblasts. Equine bone marrow stromal cells were treated with differentiation media for 18 d. A. Cells were stained with Alizarin Red on d 0 (DO) and 18 (D18). Quantification of mineralization is presented as mean ± SEM. *indicates a significant difference between d 0 and 18 (P < 0.05). Real-time reverse transcription PCR was performed on samples every 3 d (n = 6 wells/d) to determine expression of: B. markers of osteoblast differentiation [osteocalcin (OC) and alkaline phosphatase (ALP)] and C. key transcription factors [osterix (Osx), runt-related transcription factor 2 (Runx2) and T-box3 (Tbx3)]. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the endogenous control. Data are presented as fold change from d 0 (mean ± SEM). *indicates that a significant difference in mRNA expression from d 0 was observed for all genes on all d, except for Tbx3 expression on d 6 (P < 0.05).