Fig. 5

Notch activation promotes the contractile phenotype in vSMCs. a-b vSMCs-DA were transduced with AdCtrl or AdNIC for 48 h and subjected to RNA sequencing. The expression of vSMC-associated genes is shown by a heatmap (a). The enrichment score of vSMC contraction-associated genes was analyzed using GSEA (b). c-d vSMCs-DA were transduced with AdCtrl or AdNIC for 48 h. The expression of SM22α and α-SMA was determined using qRT-PCR (c) and western blotting (d) and quantitatively compared (n = 5). e AdCtrl- or AdNIC-transduced vSMCs-DA were plated in Transwell chambers and incubated for 24 h. The migration of cells to the lower chamber was evaluated by crystal violet staining and quantitatively compared (n = 5). f AdCtrl- or AdNIC-transduced vSMCs-DA were incorporated with EdU for 2 h. The percentage of EdU⁺ proliferating cells was determined under a fluorescence microscope (n = 3). g AdCtrl- or AdNIC-transduced vSMCs-DA were labeled with Dil and cocultured with bEnd.3 ECs for 2 h. After washing, vSMCs-DA adhering to ECs were determined under a fluorescence microscope and quantitatively compared (n = 5). h AdCtrl- or AdNIC-transduced vSMCs-DA cells were subjected to collagen gel contraction assay (n = 5). Relative contraction was compared. Bars = means ± SD. *, P < 0.05; **, P < 0.01; and ***, P < 0.001