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. 1998 Jul 20;188(2):351–364. doi: 10.1084/jem.188.2.351

Figure 6.

Figure 6

Figure 6

Localization of V-ATPases in early endosomes and in phagosomes. (A–C) Macrophages obtained from Nramp1 −/− mice were allowed to internalize Texas red–labeled fixable (1 mg/ml) dextran for 15 min at 37°C. The cells were then fixed, permeabilized, and incubated with an affinity-purified polyclonal antibody against the 39-kD subunit of the V-ATPase, followed by an FITC-labeled secondary antibody. (A) Localization of Texas red–labeled dextran; (B) localization of the V-ATPase subunit; (C) areas of overlap between the dextran and the ATPase. (D–F) Macrophages obtained from Nramp1 −/− mice were allowed to internalize live, fluoresceinated M. bovis and were subsequently incubated with Texas red–labeled dextran for 15 min at 37°C. The cells were then fixed and visualized by Nomarski (D) and confocal immunofluorescence microscopy. (E) Localization of fluoresceinated bacteria; (F) distribution of Texas red–labeled dextran. Images are representative of at least five experiments of each kind. (G) Macrophages obtained from Nramp1 −/− mice were allowed to internalize FITC-labeled human holotransferrin (20 μg/ml), and pHe was measured using single-cell imaging. Where indicated, concanamycin (100 nM) was added. Data represent means ± SEM of three separate experiments.