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. 2020 May 17;21(10):3539. doi: 10.3390/ijms21103539

Figure 3.

Figure 3

Secreted frizzled-related protein 5 (sFRP5) inhibited osteoblastic trans-differentiation of vascular smooth muscle cells (VSMCs) cultured in vascular calcification (VC) induction media (high-phosphate, angiotensin II, and vitamin (D). The protein level of RUNX2 was determined using Western blotting, and calcification was confirmed visually by von Kossa staining. Six replicates per condition were performed. (A) Treatment with sFRP5 of VSMCs in VC induction medium decreased the expression of RUNX2, and neutralization with anti-sFRP5 restored the expression of RUNX2 to control immunoglobulin G levels; (B) VSMCs cultured in VC induction medium with different additional interventions and stained with von Kossa stain are shown. Six replicates per condition were performed. VSMCs incubated in VC induction medium showed significantly increased staining compared with the control. Treatment with sFRP5 led to the attenuation of staining, and the addition of anti-sFRP5 resulted in increased staining. Scale bar, 100 µm. Data are expressed as means ± standard errors of the means from six independent experiments. * p < 0.05, ** p < 0.01.