Fig. 2. TCF21 promotes HASMC osteogenesis.

HASMCs were transfected with the TCF21 overexpression vector, TCF21 siRNA or the corresponding control for 24 h. The cells were then switched to complete DMEM or complete medium containing high phosphate (Pi, 3 mM) for 3 d. At the end of the experiment, the cells were used for the following assays. a, e Calcium deposition was examined by Alizarin Red S staining. *P < 0.05 (n = 4). b, f Intracellular calcium levels were determined by a calcium detection kit, and the cell protein concentration was used for relative quantitative statistical analysis. *P < 0.05 (n = 4). c The mRNA expression of TCF21, RUNX2 and BMP2 in HASMC was analyzed by qPCR. *P < 0.05 (n = 3). d, g The protein expression of TCF21, RUNX2 and BMP2 was determined by Western blotting. *P < 0.05 (n = 3).