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. Author manuscript; available in PMC: 2018 Jul 21.
Published in final edited form as: Circ Res. 2017 Jun 12;121(3):220–233. doi: 10.1161/CIRCRESAHA.116.310293

Figure 3. DRP1 inhibition attenuated human SMC osteogenic media-induced fibrocalcific response.

Figure 3

(A) Live cell intracellular calcium concentrations, and (B) TNAP mRNA, (C) protein, and (D) relative enzyme activity from human coronary smooth muscle cells (SMCs) cultured in control media (CM) or osteogenic media (OM) for two weeks with DMSO vehicle (0.01%) or Mdivi-1 (50 μmol/L). (E) Relative media pyrophosphate (PPi) from human coronary SMCs treated for two weeks. (F) Representative Alizarin Red stain image and quantification from human coronary artery SMCs treated with Mdivi-1 or DRP1 siRNA for three weeks. (G) Representative Alizarin Red stain and quantification for human aortic SMCs treated for three weeks in OM with H2O2 (0.3mmol/L), with or without Mdivi-1. (H) Human coronary artery SMC media COL1A1 ELISA from cells treated for two weeks. (I) COL1A1 mRNA from human coronary artery SMCs treated for two weeks. (J) Representative Osteosense (red; calcification) and CNA immunofluorescence (green; collagen) from human coronary artery SMCs treated for three weeks. N=3–6 donors; ns not significant, * P <0.05, **<0.01, ***<0.001, ****<0.0001; error bars indicate STDEV.