Figure 4.

The accumulation of mTOR-inhibitory protein Deptor contributed to autophagy response upon ROC1 silencing. (a and b) Cells transfected with siControl or siROC1 were subjected to IB analysis for expression of indicated proteins over time. ROC1 silencing induced Deptor accumulation (a) and mTOR inactivation, as demonstrated by notable reduction of p70S6K and p4EBP1 in HepG2 cells (b). (c–f) Autophagy was rescued by Deptor siRNA silencing. Deptor knockdown largely abrogated ROC1 silencing-induced conversion of LC3-I to LC3-II in HepG2 (c) and Huh7 (e) cells, and decreased classical punctuative distribution of EGFP-LC3 in HepG2-EGFP-LC3 (d) and Huh7-EGFP-LC3 (f) cells 96 h post transfection. The percentage of GFP-punctuate-positive cells were quantified by counting cells in 10 independent areas. The results were presented as mean value±S.E. from three independent experiments with each running in triplicate. **P<0.01