Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2013 Mar 1.

Published in final edited form as: Mol Cancer Res. 2012 Jan 24;10(3):392–400. doi: 10.1158/1541-7786.MCR-11-0359

Fig. 5 — Fig. 5A Effects of imexon and eIF2B5 silencing on protein expression. MiaPaCa-2 cells were transfected with eIF2B5 siRNA or a non-targeting sequence (si CT) and examined for protein synthesis. Data are expressed as the percent of control, where the control is untreated cells transfected with a non-targeting siRNA (Mean ± SD, n=8). Imexon significantly inhibited protein synthesis in both si CT and si eIF2B5 transfected cells (*p ≤ 0.05). Knockdown of eiF2B5 also inhibits protein synthesis and is augmented by 250 μM IMX (^p = 0.0017). There is no significant augmentation of synthesis inhibition by 500 μM IMX (p = 0.75, si elF2B5 vs. si CT).

Fig. 5B Effect of NAC supplementation on protein synthesis inhibition by imexon. MiaPaCa-2 cells were pre-treated with 10 mM NAC overnight. NAC was either maintained (“continuous”) or the NAC removed (“pre-treated only”), in the presence of varying concentrations of imexon for 24h. Alternatively, the addition of NAC was delayed until 2h after imexon exposure (“post imexon”). Protein synthesis was measured by ³H-leucine incorporation. Data are expressed as percent of control, where the control is untreated cells (Mean ± SEM, n = 20). *p<0.05, **p<0.01

Fig 5C. Effect of ROS scavengers on protein synthesis inhibition by imexon. MiaPaCa-2 cells were pre-treated for 2h with the 100 U/ml of pegylated superoxide dismutase (PEG-SOD), 200 U/ml of pegylated catalase (PEG-CAT), or 100μM of 1-oxyl-2,2,6,6-tetramethyl-4-hydroxy- piperidine (tempol) before the addition of imexon for a total of 24h. Protein synthesis was measured by ³H-leucine incorporation. Data are expressed as percent of control, where the control is untreated cells (Mean ± SEM, n = 18).

Fig. 5 — Fig. 5A Effects of imexon and eIF2B5 silencing on protein expression. MiaPaCa-2 cells were transfected with eIF2B5 siRNA or a non-targeting sequence (si CT) and examined for protein synthesis. Data are expressed as the percent of control, where the control is untreated cells transfected with a non-targeting siRNA (Mean ± SD, n=8). Imexon significantly inhibited protein synthesis in both si CT and si eIF2B5 transfected cells (*p ≤ 0.05). Knockdown of eiF2B5 also inhibits protein synthesis and is augmented by 250 μM IMX (^p = 0.0017). There is no significant augmentation of synthesis inhibition by 500 μM IMX (p = 0.75, si elF2B5 vs. si CT).

Fig. 5B Effect of NAC supplementation on protein synthesis inhibition by imexon. MiaPaCa-2 cells were pre-treated with 10 mM NAC overnight. NAC was either maintained (“continuous”) or the NAC removed (“pre-treated only”), in the presence of varying concentrations of imexon for 24h. Alternatively, the addition of NAC was delayed until 2h after imexon exposure (“post imexon”). Protein synthesis was measured by ³H-leucine incorporation. Data are expressed as percent of control, where the control is untreated cells (Mean ± SEM, n = 20). *p<0.05, **p<0.01

Fig 5C. Effect of ROS scavengers on protein synthesis inhibition by imexon. MiaPaCa-2 cells were pre-treated for 2h with the 100 U/ml of pegylated superoxide dismutase (PEG-SOD), 200 U/ml of pegylated catalase (PEG-CAT), or 100μM of 1-oxyl-2,2,6,6-tetramethyl-4-hydroxy- piperidine (tempol) before the addition of imexon for a total of 24h. Protein synthesis was measured by ³H-leucine incorporation. Data are expressed as percent of control, where the control is untreated cells (Mean ± SEM, n = 18).