Figure 8.
Knockdown of Slug attenuates warfarin-induced pAVIC calcification and rescue by p53. A, the efficacy of shRNA for knockdown of Slug was assessed by Western blotting. Relative protein levels were normalized to GAPDH. B and C, calcium deposition was assessed by Alizarin Red staining and measured with the QuantiChromTM calcium assay kit. D, ALP activity was performed to assess the amount of osteoblastic differentiation and calcification. E–H, expression of Runx2 and Bmp2 were assessed by qPCR. Expression of Slug, p53, Runx2, p-Smad1/5, and p-Akt were assessed by Western blotting. Relative protein levels were normalized to GAPDH. The data are shown as the means ± S.E. of triplicates and are representative of three independent experiments performed. Ctrl, control; War, warfarin. *, p < 0.05; **, p < 0.01; ***, p < 0.001.