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. 2013 Dec 1;24(23):3736–3745. doi: 10.1091/mbc.E13-07-0408

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© 2013 Krajcovic et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 2: — mTOR is recruited to lysosomal vacuoles harboring engulfed cells but not latex beads. (A) Top, mTOR (left) is recruited to corpse-containing entotic vacuole (arrow) (MCF10A), where it colocalizes with Lamp1 (right); inset, colocalization of immunostained mTOR (red) and Lamp1 (green). DAPI-stained nucleus is shown in blue. Bottom, mTOR does not recruit to bead-containing vacuole (left, arrow), which is marked by Lamp1 (right) (MCF10A); inset, merged image with DAPI-stained nucleus in blue. All are confocal microscopic images. Bars, 10 μm. (B) mTOR localizes to corpse-containing entotic vacuole (labeled corpse with arrow; 70% were positive for mTOR, n = 37) but not to bead-containing lysosomal vacuole (bead, arrow; 2.3% positive for mTOR, n = 84) in the same MCF10A cell. Top and middle, confocal images of immunofluorescence for mTOR and Lamp1; bottom, merge with DAPI-stained nucleus (blue); inset, DIC. (C) mTOR localizes to apoptotic cell phagosomes (52% positive for mTOR, n = 54) but not latex bead phagosomes (1.9% positive for mTOR, n = 154) in J774.1 macrophages. Confocal microscopic images show macrophage with an engulfed apoptotic corpse and two beads, as indicated, stained for mTOR and Lamp1 by immunofluorescence. Right, merged image with DAPI-stained nucleus (blue) and DIC.