FIG. 6.

siRNA-mediated knockdown of Jagged1 reduces the expression levels of SMC-specific marker proteins associated with downregulation of Notch3 in 3D coculture. HCASMCs were seeded on 3D scaffolds and were cultured prior to coculture. HCAECs were transiently transfected with Jagged1 siRNA or scrambled siRNA (as a control), seeded on the HCASMC layer, and cocultured for 2 days. Following separation of cells, Western blot analysis revealed that transfection was successful and the corresponding Jagged1 expression in HCAECs was significantly reduced (A). Notch3 was significantly downregulated in HCASMCs by siRNA-mediated knockdown of Jagged1 as determined by western blot analysis (B). Both SM-α-actin and calponin were significantly downregulated, suggesting that Notch3 signaling is responsible for contractile VSMC phenotype regulation in 3D cultures (C, D). Data are represented as mean±SD from three independent experiments (*p<0.05).