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. 2017 Nov 22;59(1):69–78. doi: 10.1194/jlr.M079731

Fig. 3.

Fig. 3.

The effect of C2-ceramide and S1P on VSMC matrix mineralization. A: VSMCs were cultured in osteogenic (Osteo) or osteogenic with 10 μM C2-ceramide medium. At mid mineralization, the cells were stained with alizarin red. Representative photomicrographs showing mineralization (red), scale bar = 500 μm [A(i)] and quantification of matrix mineralization by elution of alizarin red [A(ii)]. Data are mean ± SEM from six independent experiments. *P < 0.05 osteogenic versus osteogenic plus C2-ceramide by paired t-test. B: VSMCs were cultured in osteogenic medium with the indicated concentrations of S1P. At early mineralization, the cells were stained with alizarin red. Representative photomicrographs showing mineralization (red), scale bar = 500 μm [B(i)] and quantification of matrix mineralization by elution of alizarin red [B(ii)]. Data are mean ± SEM from three independent experiments. *P < 0.05 osteogenic versus osteogenic plus S1P by one-way ANOVA with Tukey’s multiple comparisons post test.