FIGURE 4.

NOTCH3 expression in human dental pulp mesenchymal stem cells (MSCs) in vitro. A, Immunofluorescent staining showing distribution of NOTCH3 (green color), JAG1 (red color), Phalloidin (white color) and DAPI (blue color) in cultured dental pulp MSCs. B‐D, Staining showing multilineage differentiation potential of human dental pulp MSCs. Differentiation was obtained by culturing human dental pulp cells in osteogenic, adipogenic, and chondrogenic media for 3 weeks. B, Alizarin Red staining performed on dental pulp MSCs cultured in osteogenic conditions; C, Oil Red O staining performed on dental pulp MSCs cultured in adipogenic conditions; D, Alcian Blue staining performed on dental pulp MSCs cultured in chondrogenic conditions. E, Real‐time PCR analysis of the expression of NOTCH3 in dental pulp MSCs undergoing osteogenic/odontoblastic differentiation. F, Real‐time PCR analysis of the expression of the osteogenic and odontoblastic differentiation markers DSPP (Dentin Sialophosphoprotein), ALP (Alkaline Phosphatase), and OSX/SP7 (Osterix) in dental pulp MSCs undergoing osteogenic/odontoblastic differentiation. **P‐value <.01; *P‐value <.05. N = 4 for each time point and condition. Error bars indicate standard deviation. Scale bars = A = 25 μm; B = 100 μm; C = 50 μm; D = 200 μm