Figure 4.

The PDCD4 levels were upregulated by the autophagy inhibitors bafilomycin A1 and 3-MA and by the proteasome inhibitor MG132 in ATG5 mutant cells. (A) After reaching 90–95% confluency, the ATG5 mutant Huh7 cells were treated with DMSO (D) as a control, 10 µM bafilomycin A1 (B), 0.1 nM rapamycin (R) and 20 µM MG132 in the presence or absence of EGF in normal or serum-free medium and incubated for 4 h. The cells were collected and Western blotting analyses were performed by using antibodies against the components mentioned in the figure. (B) ATG5 mutant Huh7 cells at 90–95% confluency were treated with 5 mM 3-MA and the culture continued for 6, 9, and 12 h, after that cells were collected for Western blotting. Experiments were repeated at least three times and similar results were obtained. The representative results are demonstrated.