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. 2022 Feb 7;10:774363. doi: 10.3389/fcell.2022.774363

FIGURE 3.

FIGURE 3

miR-140-5p antagonises osteogenic differentiation of VSMCs. (A) qRT-PCR was performed to evaluate the expression of miR-140-5p in VSMCs cultured in OM medium. (B) qRT-PCR was performed to evaluate the expression of miR-140-5p in VSMCs transfected with specific miR-140-5p mimics or inhibitor. (C) Western blotting was performed to determine the protein expression of Runx2 and ALP in VSMCs transfected with specific miR-140-5p mimics or inhibitor. The data were presented as densitometric ratios normalized to GAPDH [(C), lower panel]. (D) qRT-PCR was performed to determine the protein expression of Runx2 and ALP in VSMCs transfected with specific miR-140-5p mimics or inhibitor. (E) The ALP activity was evaluated by specific Elisa kits in VSMCs transfected with specific miR-140-5p mimics or inhibitor. (F) Alizarin Red S staining were measured in ß-GP-treated VSMCs transfected with specific miR-140-5p mimics or inhibitor for 21 days. Representative microscopic views are shown. Scale bar represents 200 µm. Data were presented as ratio of positive staining area [(F), right panel]. Each experiment was repeated for three times. The data represent the mean ± SD of triplicates. *p < 0.05.