Dexamethasone Regulates EphA5, a Potential Inhibitory Factor with Osteogenic Capability of Human Bone Marrow Stromal Cells

Figure 1

Forced expression of EphA5 inhibits osteoblast differentiation in hBMSCs. (a) Lentiviral constructs for transducing Luc and EphA5. (b) Schematic representation of the cell culture protocol. Adult hBMSCs were transduced with LV-Luc or LV-EphA5. ((c)–(f)) Quantitative analysis of mRNA expression of osteogenic markers at 7 days of osteogenic culture: (c) EphA5, (d) ALP, (e) Runx-2, and (f) ITGA5 (). The fold change of gene expression was normalized against the expression in cell cultures without LV transduction. ((c), lower) EphA5 protein levels were determined by western blot analysis. (g) Adhesion assays (). Images of cell attachment (upper). ((h)–(j)) CFU assays (). (h) Number of total colonies. (i) CFU-ALP positive rate (upper) and number of ALP-positive colonies (lower). Images of wells after ALP staining (middle). (j) CFU-OB positive rate (upper) and number of OB-positive colonies (lower). Images of wells after von Kossa staining (middle).

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