Figure 6.

Autophagy inhibition leads to AOPP-BSA-induced endothelial senescence. (A) Effects of autophagy activation with Rapa and inhibition with 3-MA on AOPP-BSA-induced expression of senescence-associated protein p21 and p16 were measured by western blot. HUVECs were pretreated with Rapa (200 nM) or 3-MA (5 mM) for 2 h before AOPP-BSA stimulation. GAPDH was used as loading control. (B,C) Quantification of p21 and p16 protein expression (n = 4). (D) HUVECs were pretreated as mentioned above. Effects of Rapa and 3-MA on HUVEC senescence were also detected by SA-β-gal assay. Scale bar for upper panel = 20 μm, scale bar for lower panel = 50 μm. (E) The SA-β-gal positive rate was quantitated (n = 4). All data analyzed by one-way ANOVA with LSD or Dunnett's T3 post-hoc multiple comparisons are shown as mean ± SEM. Rapa, rapamycin; 3-MA, 3-methyladenine.