Figure 4. Effects on VSMC calcification induced by inhibiting PFKFB3 expression by β-GP. (A) Representative blot images of osteogenic differentiation-associated proteins. (B) Quantitative analysis of RUN2 protein expression using densitometry. (C) Quantitative analysis of BMP2 protein expression using densitometry. (D) Quantitative analysis of SM22a protein expression using densitometry. (E) Calcium nodules were stained with Alizarin red. Red nodules indicate calcium deposition. Scale bar = 50μm. (F) Quantification of mineralization. β-GP, β-Glycerophosphate disodium salt pentahydrate; Data were normalized using log10 and analyzed using one-way ANOVA tests; U, U50,488H; 3-PO, a novel small molecule inhibitor of the PFKFB3 isozyme; Data obtained from quantitative densitometry were presented as means ± SEM. n=5 in each group. **P < 0.01 versus the control group, #P < 0.05 versus the β-GP group, ##P < 0.01 versus the β-GP group.