Fig. 2. Zingerone inhibits Pi-induced calcification of VSMCs. VSMCs were cultured with inorganic phosphate and/or zingerone for 2 days or 2 weeks. (A-C) mRNA expression of Dlx5, CBFA1 and ALP was measured by real-time polymerase chain reaction (A). Protein levels of Dlx5, CBFA1 were measured by Western blot analysis (B, left panel). Densitometry analysis was performed with the indicated antibodies (B, right panel). Alizarin red S staining (C) Data represent the mean±standard error of the mean of 3 individual experiments.

Pi, inorganic phosphate; VSMC, vascular smooth muscle cell; Dlx5, distal-less homeobox 5; CBFA1, core-binding factor α-1; ALP, alkaline phosphatase.

Statistical significance was determined relative to the control (^*p<0.05; ^†p<0.01; ^‡p<0.001) or compared with Pi-treated cells (^§p<0.05; ^∥p<0.01) using the Student's t-test.