Figure 2
Inhibition of PKCĪ“ activity attenuates osteogenic differentiation in hBMSCs. (A) Post-confluent hBMSCs were cultured in GM or ODM at the indicated concentrations of rottlerin, a specific PKCĪ“ inhibitor. ALP activity was determined after osteogenic induction of hBMSCs for 7 days. (B) For quantitative determination of ALP activity, the absorbance at 405 nm of cell extracts was measured and expressed as the fold change of treated cells over vehicle-treated cells in GM. (C) Post-confluent hBMSCs were cultured in GM or ODM with 2 Ī¼M rottlerin. Alizarin red S staining was performed to monitor mineralization of hBMSCs after osteogenic induction for 14 days. (D) For quantitative determination, the absorbance of alizarin red S was measured at 570 nm and expressed as the fold change of treated cells over vehicle-treated cells in GM. (E) Post-confluent hBMSCs were cultured in GM or ODM with 2 Ī¼M rottlerin, and then harvested at 14 days. The mRNA expression of osteogenesis-related genes, including ALP, RUNX2, and OCN, was estimated by RT-PCR. The representative images from three independent experiments are shown. Data shown are means Ā± S.E. (*P <0.05 versus vehicle-treated cells in ODM) of three independent experiments.