Figure 3.

Bavachin inhibits β-GP-induced calcification in HASMCs. (A) Mineralization of calcium was quantified by VON KOSSA staining in bavachin and β-GP-treated HASMCs. The blackspots in the image are calcified deposition areas; magnification 10×. (B) Intracellular Ca²⁺was quantified by FLIPR calcium 6 dye (green) in bavachin- and β-GP-treated HASMCs; magnification 4×. (C) Flow cytometry analysis of intracellular Ca²⁺level in bavachin and β-GP-treated HASMCs. Statistical significance was analyzed by the t-test (**P < 0.01, ***P < 0.001). (D) Western blot analysis of calcification-related proteins (Wnt3a, β-catenin, OPN, BMP2, OPG, and Runx2) in HASMCs. The full-length images of Western blot are shown in Figure S3D . Statistical significance was analyzed by the one-way ANOVA (*P < 0.05, **P < 0.01, ***P < 0.001). The data is represented as mean ± S.D. (n = 3).