FIGURE 4.

Lysosome inhibitors can inhibit MTORC1 and initiate autophagosome formation. A, MEFs were treated with Baf (0.5 μm), ConA (2.5 μm), AC (40 mm), CQ (40 μm), or E64D (25 μm) plus pepstatin A (50 μm) (E/P) for 0 to 22 h, followed by immunoblot analysis. B and C, A549 cells were treated as indicated with or without 3-MA (10 mm) for 6 h, followed by immunostaining with anti-ATG16L1 (B) or anti-ATG12 (C). D and E, A549 cells were treated as indicated (40 mm AC; 1 μm rapamycin; 10 mm 3-MA) for 6 h, followed by immunostaining with anti-ATG16L1 (D) or anti-ATG12 (E), which were quantified. For panels D and E, values represent means ± S.D. from three independent experiments. **, p < 0.01; *, p < 0.05. DMSO, dimethyl sulfoxide; CM, complete medium; Rap, rapamycin.