Fig. 3

Increase of autophagy of ADR-treated cells in HG was in part depended on p53 activity. (a) HCT116 cells were treated with ADR (2 μg/ml) for 16 h in LG and HG medium, with or without p53 inhibitor pifithrin-α (PFT-α) (30 μM) before being assayed for western blot analysis. Densitometric values of LC3II/I/β-actin protein levels were quantified using the ImageJ software and normalized to control. One representative experiment is shown. (b) HCT116 cells were transfected with ctr-siRNA and siDRAM and 36 h after transfection DRAM expression was assessed by RT-PCR analysis. One representative experiment is shown. (c) HCT116 cells, transfected with ctr-siRNA and siDRAM, were kept in HG medium for 24 h and then treated with ADR (2 μg/ml) for 16 h before the expression of p62 was measured by western blot analysis. Densitometric values of p62/β-actin protein levels were quantified using the ImageJ software and normalized to control and the results are shown under the images. One representative experiment is shown. Anti β-actin was used as protein loading control