Upregulation of miR-664a-3p Ameliorates Calcific Aortic Valve Disease by Inhibiting the BMP2 Signaling Pathway

Figure 2

miR-664a-3p inhibits osteogenic differentiation and calcification of valvular interstitial cells. (a) RNA-FISH revealed the localization of miR-664a-3p in VICs. miR-664a-3p is stained in red and the nucleus in blue. Scale bar, 20 μm. (b) RT-qPCR was conducted to detect the miR-664a-3p expression in VICs after overexpression with a mimic or knockdown by an inhibitor. (c) Western blotting analysis of RUNX2, MST2, and OSTERIX expression in VICs after alterations in the level of miR-664a-3p. Quantitative analysis of the protein bands are shown on the right. (d) The formation of calcium nodules after alterations in the level of miR-664a-3p was detected by Alizarin Red staining. Quantitative analysis is shown on the right. Scale bar, 50 μm. (e) ALP activity in VICs after alterations in the level of miR-664a-3p was detected and quantitated by ALP staining assay. Scale bar, 200 μm. , , and .

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