Figure 2.

The effects of warfarin on p53 expression. A, p53 mRNA levels with and without warfarin treatment were assessed by qPCR. B, protein levels of p53 and phosphorylation of p53 were assessed by Western blotting. C, acetylated p53 were immunoprecipitated by Ac-lysine antibody and blotted with p53 antibody. D, relative protein levels of p53, p-p53, and Ac-p53 were normalized to GAPDH, p53, and Ac-lysine. E, representative images of Alizarin Red staining of mouse aortic valve. F, p53 expression levels in calcific and control mouse aortic valves were assessed by qPCR (n = 5 for each group). G, representative images of immunofluorescent staining of mouse aortic valves with the indicated antibodies. Scale bar, 50 μm. *, p < 0.05; Ctrl, control; War, warfarin; IP, immunoprecipitation; IB, immunoblot; Vim, vimentine.