Fig 6.

Umb did not stimulate osteoblastic differentiation. (A) Primary osteoblasts were cultured in the presence of AA (50 μg/mL), β-GP (10 mM), and different amounts of Umb for 6 days during osteoblastic differentiation. ALP positive cells were monitored by ALP staining (upper). ALP activity is expressed as p-nitrophenol released (lower). (B) Calcium deposits for matrix mineralization were measured by Alizarin Red S (ARS) staining after 21 days (upper). The intensity of staining was quantified with cetylpyridinium chloride (lower). (C) The mRNA expression levels of Runx2, ALP, COL1A1, and BSP were analyzed by real-time RT-qPCR.