Fig. 4From: p53-Dependent PUMA to DRAM antagonistic interplay as a key molecular switch in cell-fate decision in normal/high glucose conditionsAutophagy inhibition or DRAM silencing restored ADR-induced cell death in HG. (a) HCT116 cells were treated with ADR (2 μg/ml for 24 h) in low (−) and high glucose (HG) condition with or without 25 μM CQ (for 16 h). After treatments, cells were in part fixed and stained with propidium iodide (PI) for subG1 evaluation (upper panel) or lysed and analyzed by western immunoblotting to assess PARP cleavage (lower panel); relative quantification of PARP cleavage/β-actin ratio is shown. One representative experiment is shown. Anti-β-actin was used as protein loading control. *P < 0.001. (b) RKO and HCT116 cells, transfected with ctr-siRNA and siDRAM or left untransfected, were kept in low glucose (−) or high glucose (HG) medium for 24 h and then treated with ADR (2 μg/ml) for 24 h before the percentage of dead cells was scored by trypan blue exclusion. The data are presented as the means ± S.D. from three independent experiments. *P < 0.001. (c) RKO cells, transfected with ctr-siRNA and with siATG5 or left untransfected, were kept in low glucose (−) or high glucose (HG) medium for 24 h and then treated with ADR (2 μg/ml) for 24 h before the percentage of dead cells was scored by trypan blue exclusion. The data are presented as the means ± S.D. from three independent experiments. *P < 0.001Back to article page