Fig. 5

Autophagy inhibition or DRAM silencing restored ADR-induced PUMA transcription in HG. (a) RKO and HCT116 cells were kept in low glucose (LG) or high glucose (HG) medium for 24 h and then treated with ADR (2 μg/ml) for 16 h in the presence or absence of 25 μM CQ, before being assayed for semi-quantitative RT-PCR analysis of PUMA gene expression. 28S was used as a control for efficiency of RNA extraction and transcription. One representative experiment is shown. (b) RKO and HCT116 cells, transfected with with siRNA and siDRAM or untransfected, were kept in low glucose (−) or high glucose (HG) medium for 24 h and then treated with ADR (2 μg/ml) for 16 h before being assayed as in (a). One representative experiment is shown. (c) Densitometric values of three independent experiments as in (b) were quantified using the ImageJ software and normalized to control. The values of control were set to 1. The data are presented as fold of induction ± S.D. *P < 0.005