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

Fig. 6.

Fig. 6.

Inhibition of S1P-dependent activation of ERM prevents VSMC matrix mineralization. A: VSMCs were incubated with vehicle (0.1% DMSO), 1 μM NSC668394 (NSC), or 10 μM NSC668394 for 16 h before stimulation with 10 μM S1P for 10 min, and cell lysates prepared for immunoblot analysis with pERM. β-Actin was used as a loading control (Con). Representative immunoblot and densitometric data of pERM corrected for loading are shown. The data are mean ± SEM from three independent experiments. *P < 0.05 for control versus 10 μM S1P by two-way ANOVA with Dunnett’s multiple comparisons post test. B: VSMCs were cultured in osteogenic (Osteo) medium, osteo plus NSC668394 (NSC) (10 μM), osteo plus S1P (10 μM), or osteo plus NSC668394 (10 μM) plus S1P (10 μM). At the mid-mineralization time point, cells were stained with alizarin red. Representative photomicrographs showing mineralization (red), scale bar = 500 μm and quantification of matrix mineralization by elution of alizarin red. Data are mean ± SEM from six independent experiments. *P < 0.05 by one-way ANOVA with Tukey’s multiple comparisons post test.