Fig. 4: Effect of Met following TERT knockdown during the OA-induced pathogenesis of atherosclerosis and on senescent phenotypes.

a VSMCs were transfected with control siRNA or TERT siRNA (10 μM) for 48 h, pretreated with 200 μM OA for 24 h, and incubated with 2 mM Met for 1 h, after which the protein levels were analyzed by Western blotting. b Cellular senescence was assessed by SA-β-gal staining and bright-field microscopy (200x, scale bar = 100 μm). c After transfection, the protein levels were analyzed by Western blotting with TNF-α and ADRP antibodies and d the cells were stained with BODIPY493/503 (a green lipid stain) (scale bar = 30 μm). e After transfection, colocalization of TERT (green) and 53BP1 foci (red) was observed by confocal microscopy. Nuclei were stained with DAPI (blue). Magnified views of merged images showing details of the colocalization are shown in the lower series of panels (scale bar = 30 μm). f Western blotting analysis of contractile (elastin and α−SMA) and synthetic proteins (collagen I) in VSMCs. Representative results are shown, and the values are the means ± SEs of three independent experiments.