Fusion and remodeling of <i>L. amazonensis</i> PVs.

<p>(A) Fusion between <i>L. amazonensis</i> PVs recorded by time-lapse microscopy of infected macrophage cultures loaded with Lysotracker (green). Asterisks indicate PVs involved in fusion; arrowheads indicate an intermediary PV morphology following membrane fusion. Time-lapse acquisition started after 48 h of infection, and the time of acquisition is shown as h:mm. Bar = 10 µm. (B–C) Multidimensional imaging of <i>L. amazonensis</i> PVs loaded with Lysotracker in live infected macrophages. In B, Lysotracker fluorescence of an infected macrophage containing four PVs. Asterisks indicate the PVs involved in fusion. In C, the isosurfaces representative of each PV in the multidimensional image. Isosurfaces have statistic-based color according to their measured volumes ranging from cyan (smaller volume) to magenta (larger volume). Fusion-prone PVs and fused vacuole are indicated. Image acquisition started after 48 h of infection, and the time of acquisition is shown as d:hh:mm. Bar = 10 µm. (D) Volume measurements of <i>L. amazonensis</i> PVs (a–d) in an infected macrophage (data acquisition started from 2 h of infection). Fusion between PVs is indicated by arrowheads, and the PVs involved in fusion are identified (i.e., when b fused with d, b–d). (E) Volume measurements of <i>L. amazonensis</i> PVs (a–d) in two different infected macrophages (data acquisition started from 48 h of infection). Fusion between PVs is indicated by arrowheads, and the PVs involved in fusion are identified. Another 5 multidimensional images of fusion-prone PVs were also analyzed and presented the same results.