Figure 6.

Effect of uL3 status on the conversion of LC3B and on the expression of autophagy-related genes. Representative immunoblotting showing LC3B protein conversion (A) and p62 protein levels (C) in HCT 116^p53−/−cells and LC3B protein conversion (B) and p62 protein levels (D) in uL3ΔHCT 116^p53−/− cells treated or not with 25 µM CQ or transiently transfected with 1 µg of pHA-uL3. 24 h later, protein extracts from the samples were analyzed by Western blotting with antibodies against indicated proteins. GAPDH and β-actin were used as loading controls. Quantification of LC3B-II/I ratio is shown. Full-length blots are shown in Figures S7 and S8. (E) Total RNA from HCT 116^p53−/− and uL3ΔHCT 116^p53−/− cells, transfected or not with 1 µg of pHA-uL3, was subjected to RT-qPCR with primers specific for the indicated genes (Table 1). Quantification of signals is shown. Bars represent the mean of triplicate experiments; error bars represent the standard deviation. * p < 0.05, ** p < 0.01 vs. untreated cells set at 1.