Fig. 5

A, B Light absorbance at 340 nm reveals that EVs from 10% stretched VSMCs form more calcific mineral. C Normalization with respect to control samples reveals higher mineralization rate for 10% stretch. D Annexin V, a calcium binding membrane protein, in EVs increased in response to increases in mechanical stretch. Collagen hydrogel assay to visualize EV-induced mineralization for stretched cells (E) compared to non-stretched control (F). G Mechanical stretch significantly elevated the number of the mineral nucleation sites. H) Total mineralized area was increased by EVs from mechanically stretched VSMCs. n ≥ 4. Data are mean ± SD (*P < 0.05, **P < 0.01). (Scale bar: 10 µm). Full-length blot is presented in Supplementary Fig. 5