Targeting lysosomal degradation induces p53-dependent cell death and prevents cancer in mouse models of lymphomagenesis
J. Clin. Invest. Kirsteen H. Maclean, et al. 118:79 doi:10.1172/JCI33700 [Go to this article.]

Figure 5
CQ provokes markers of macroautophagy yet inhibits lysosomal functions. (A) CQ induces the accumulation of PE-modified LC3. Early passage (p2) MEFs were treated with 50 μM CQ for 24 h. Expression and modification of LC3 (pro-form, LC3-I, and PE-modified LC3-II) was analyzed by western blot. (B) Indicated MEFs were treated for 24 h with 4-HT to activate Myc-ER^TAM and with or without 50 μM CQ for 24 h. Expression and modification of LC3 were monitored by western blot analyses. (C) Top: GFP-LC3B–expressing MEFs were incubated with 100 nM Lysotracker for 30 min with or without CQ (50 μM) for 6 h. Cells were then imaged using a Nikon inverted confocal fluorescent microscope. Bottom: Time course analyses of CQ-induced changes in GFP-LC3B–expressing MEFs. Cells were incubated with 100 nM Lysotracker for 30 min, then treated with CQ (50 μM), followed by real-time video microscopy (see Supplemental Videos 1 and 2). Images were taken at the indicated times using a Nikon inverted confocal fluorescent microscope. Original magnification, ×63. (D) Wild-type early passage (p2) MEFs were treated with or without CQ (50 μM) for 4 h or 24 h. Cells were fixed with 2% (vol/vol) glutaraldehyde, and 1-μM sections were analyzed by transmission electron microscopy. Magnification, ×5,000. Scale bars: 1 μM. L, lysosome; A, autophagosome; AL, autophagolysosome. (E) CQ induces the accumulation of p62 and cathepsin D. MEFs were treated for 24 h with 4-HT to activate Myc-ER^TAM and then treated with or without CQ for 24 h. Expression of p62 and cathepsin D was analyzed by western blot. *NS, nonspecific.