Fig. 3.

Repression of mTORC2 formation alleviated endothelial senescence. a HUVECs were transfected with different concentrations of Rictor siRNA or the negative control (NC, nonspecific siRNA) for 48 h; then, cells were harvested for Western blotting analysis for the detection of Rictor, n = 3. b HUVECs transfected with 100 nM Rictor siRNA were harvested for Western blotting analysis for the detection of mTOR and p-mTOR-S2481, n = 3. c HUVECs carrying NC siRNA or Rictor siRNA were grown with or without H₂O₂, and the p-Akt and Akt protein levels were detected, n = 3. d HUVECs transfected with different concentrations of Rictor siRNA were incubated with or without H₂O₂ and then subjected to β-gal staining, n = 3. e Replicative senescent HUVECs transfected with different concentrations of Rictor siRNA were subjected to β-gal staining, n = 3. f HUVECs carrying NC siRNA or Rictor siRNA were grown with or without H₂O₂, and the p53 and p21 protein levels were detected, n = 3–6. g Young and replicative senescent HUVECs were transfected with Rictor siRNA, and the p53 and p21 protein levels were detected, n = 3. β-Actin served as the loading control. The bar graphs show the expression levels. Data are shown as the means ± S.E.M. *P < 0.05, **P < 0.01, ***P < 0.001 vs. NC group or young group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs. NC plus H₂O₂ group or NC plus senescent group