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. 2000 Nov 6;192(9):1261–1272. doi: 10.1084/jem.192.9.1261

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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graphic file with name JEM000821.f2a.jpg — The acquisition of fluid-phase lysosomal markers by L. pneumophila vacuoles. Macrophages whose lysosomes were prelabeled by endocytosis of TR-OV or TR-DEX were infected with L. pneumophila for the period indicated, fixed, incubated with DAPI to label the bacteria, then analyzed microscopically. (A) At 8 h, bacterial vacuoles were isolated from the endocytic network (black arrowheads); by 16 h, the lumen of many vacuoles harboring L. pneumophila contained a fluorescent probe (white arrowheads). The 24-h panel reveals two separate vacuoles heavily laden with bacteria; the one at left had obviously fused (white arrowheads), whereas TR-OV accumulation by the vacuole at right was below the limit of detection (black arrowheads). (B) The percentage of 50 vacuoles that contained either TR-OV (filled symbols) or TR-DEX (open symbols) at the time indicated was scored by fluorescence microscopy in four separate experiments, each represented by a different symbol (diamonds or squares).

graphic file with name JEM000821.f2b.jpg — The acquisition of fluid-phase lysosomal markers by L. pneumophila vacuoles. Macrophages whose lysosomes were prelabeled by endocytosis of TR-OV or TR-DEX were infected with L. pneumophila for the period indicated, fixed, incubated with DAPI to label the bacteria, then analyzed microscopically. (A) At 8 h, bacterial vacuoles were isolated from the endocytic network (black arrowheads); by 16 h, the lumen of many vacuoles harboring L. pneumophila contained a fluorescent probe (white arrowheads). The 24-h panel reveals two separate vacuoles heavily laden with bacteria; the one at left had obviously fused (white arrowheads), whereas TR-OV accumulation by the vacuole at right was below the limit of detection (black arrowheads). (B) The percentage of 50 vacuoles that contained either TR-OV (filled symbols) or TR-DEX (open symbols) at the time indicated was scored by fluorescence microscopy in four separate experiments, each represented by a different symbol (diamonds or squares).