Research Paper Volume 2, Issue 8 pp 487—503

Figure 6. Role of mTOR in hyperglycemic damage of HUVEC cells. (A) a Effect of glucose, Rapamycin and mTOR knock-down on survival of growth factor-starved HUVECs. Values are relative to cell survival in high glucose (10-15% survival). Numbers are mean± SD of four samples from two independent experiments. All the indicated comparisons were significant by at least p<0.05 (two-tailed unpaired T-test). b Representative Forward/Side scatter plots of live (Region R2) and dead (Region R1) cells under high glucose and high glucose + Rapamycin. Raw numbers indicate percentages with respect to all the plotted events, including cell debris. Survivals were calculated on relevant regions only, according to the formula %survival= %R2/(%R1+%R2). (B) Western blot analysis of GlcNAcylated proteins in total lysates of HUVEC cells. Glucose, Rapamycin and mTOR knock-down were combined as indicated. Impact of treatments on mTOR signaling was evaluated by anti phospho S6 immunoblotting (b). Equal protein loading was verified by anti-actin staining. Blots representative of two independent experiments.