FIGURE 6.

3-MA disrupts the function of mTOR complex I. A, 3-MA suppresses the interaction between ULK1 and mTOR complex 1. HEK293T cells were transfected with FLAG-ULK1 for 36 h, and cells were then treated in EBSS or with 3-MA (5 mm), wortmannin (50 nm) in full medium for 1 h (left) or 9 h (right). The cell lysate was subjected to immunoprecipitation (IP) using an antibody against FLAG, followed by immunoblotting. A fraction of the cell lysate was used for immunoblotting as indicated. B, the effect of 3-MA on autophagy in Tsc2^−/− MEFs. WT and Tsc2^−/− MEFs were treated with 3-MA (5 mm) or rapamycin (10 nm) for 9 h, and the cell lysate was subject to immunoblotting.