Fig. 3.

RTEF-1 prevents β-GP-induced calcification and osteogenic differentiation of human VSMCs. RTEF-1-overexpressed VSMCs, RTEF-1 siRNA VSMCs, or control VSMCs were treated in the presence of 10 mM β-GP for 9 days (n = 3). a Calcium deposition in cultured VSMCs was detected by alizarin red S staining. Scale bar = 80 mm. b Calcium content was measured. c ALP activity was assessed by spectrophotometry. d Osteocalcin mRNA expression was determined by qRT-PCR. e Runx2 mRNA expression was analyzed by qRT-PCR. f α-SMA mRNA expression was detected by qRT-PCR. g Western blot analysis of Osteocalcin, Runx2, and α-SMA protein expressions. Statistical significance was assessed using t-test. All graphs show mean + SD. *P < 0.05 vs. Control. ^#P < 0.05 vs. NC siRNA