Fig. 9

T007 promotes BMSCs differentiation into osteoblasts in vitro and expression of the osteoblast-specific genes. a ALP expression in BMSCs cultured in the osteogenic medium and treated with various concentrations of T007 for 7 days. Mineralized extracellular matrix was detected using Alizarin Red S staining for 21 days. b Expression of the osteoblast-specific genes Runx2, OCN, ALP, and COL1a in BMSCs with T007-treated maintained in osteogenic medium for 7 days. c The number of ALP-positive BMSCs and the OD values obtained for mineralized matrix solutions following the treatment with T007. d, e BMSCs were treated with various concentrations of T007, and the expression of the indicated proteins was detected by western blotting. f ALP expression in BMSCs and mineralized extracellular matrix was detected using Alizarin Red S staining treated following the transfection with pcDNA-PPARγ or the empty vector for 7 days and 21 days. g, h The number of ALP-positive cells following the transfection with pcDNA-PPARγ or the empty vector for 7 days. i, j BMSCs were transfected with pcDNA-PPARγ or the empty vector. Cell lysates were analyzed using western blotting. The expression of Runx2, PPARγ was evaluated. k Expression of the osteoblast-related genes Runx2, OCN, ALP, and COL1a in BMSCs following the transfection with pcDNA-PPARγ or the empty vector. l ALP expression in BMSCs cultured in the osteogenic medium and mineralized extracellular matrix was detected using Alizarin Red S staining treated after transfected with two kinds of siRNA or the negative control for 7 days and 21 days. m The number of ALP-positive cells and the OD values obtained for mineralized matrix solutions following transfected with siRNA or the negative control. n, o BMSCs were transfected with two kinds of siRNA or the negative control. Cell lysates were analyzed using western blotting. The expression of PPARγ, Runx2 was evaluated. p-s The expression levels of the osteoblast-specific genes (Runx2, OCN, OSX, ALP, and COL1a) in BMSCs following the transfection with two kinds of siRNA or the negative control. t BMSCs were treated with or without 0.48 μM of T007 for the indicated periods. Cell lysates were analyzed using western blotting. The expression of PPARγ was evaluated. u BMSCs were treated with T007 (0.48 μM) or MG 132 (1 μM) for 7 days, and the expression of PPARγ was detected by western blotting. All experiments were performed at least three times. *P < 0.05 and **P < 0.01 compared with the control group