Figure 1

Detection of SLUG expression by quantitative RT-PCR. The level of SLUG, RUNX2 Lef-1, SNAIL1 and SNAIL3 expression was examined by quantitative RT-PCR in three hMSC samples cultured up to 28 days in osteogenic medium (A) and in five hOB samples (B). The cDNA obtained from total RNA was subjected to quantitative TaqMan RT-PCR for SLUG, RUNX2, Lef-1, SNAIL1 and SNAIL3 transcript analysis. The experiments were carried out in triplicate, the expression levels were normalized on the basis of GAPDH expression and results of the experiments are reported as relative mRNA expression levels. ΔΔCt method was used to value the gene expression; standard error of the mean (SEM) was calculated. The commitment to osteoblastic lineage of hMSCs was evaluated by Alizarin Red staining for extracellular calcium deposition. The authentic osteoblast phenotype was confirmed in hOBs by staining for alkaline phosphatase (ALP) activity and mineralized matrix deposition (AR, Alizarin Red staining). * = p < 0.05 (respect to day 0).