Oxidative Stress Induces Endothelial Cell Senescence via Downregulation of Sirt6
Sirt6 overexpression attenuates endothelial cell senescence induced by H2O2. ECs were infected with adenovirus carrying Sirt6 gene (Ad-Sirt6) or empty vector (control, Ad-Con) and treated with 75 M H2O2 or vehicle (H2O) for 4 days. (a) SA--gal staining was performed using a Senescence Detection kit. Images were taken under a bright-field microscope at a magnification of 100x. Arrows show SA-β-gal positive cells (blue cells). The percentage of SA--gal positive cells versus total cells was determined and quantitative data were shown in the bar graph. (b) Cells were labeled with BrdU followed by staining with DAPI (blue) for nuclei and anti-BrdU antibody (Green). Images were taken at 100x magnification. Arrows show BrdU positive nuclei (green). (c) BrdU positive nuclei (green) and total nuclei (blue) were counted. The percentage of BrdU positive nuclei versus total cell nuclei was calculated and quantitative data were shown in the bar graph. (d) The cell number in each treatment was counted at different times. compared with cells infected with Ad-Con and treated with vehicle. compared with cells infected with Ad-Con and treated with H2O2.