File:The-Diverse-and-Dynamic-Nature-of-Leishmania-Parasitophorous-Vacuoles-Studied-by-Multidimensional-pntd.0001518.s006.ogv

English: Fission of L. major PVs. (A) Fission inferred by differences in Lysotracker RFIs surrounding dividing parasites. Multidimensional imaging of dividing L. major-DsRed2 amastigotes (red) hosted by a macrophage loaded with Lysotracker (green). Isosurfaces were attributed to replicating amastigotes, allowing the measurement of Lysotracker RFI surrounding parasites. The amastigotes remain next to each other after replication and the vacuolar interface between them is observed at the initial time points. An increase in the Lysotracker signal was detected in one dividing amastigote, whereas the other remained in a PV with a low Lysotracker signal. By the end of the recordings, the amastigotes were surrounded by Lysotracker at low intensities, and no Lysotracker-positive vacuolar interface was observed between them. Image acquisition started after 48 h of infection, and the time of acquisition is shown as hh:mm. (B) L. major PV fission observed in infected RAW 264.7 macrophages expressing LAMP1 and Rab7 tagged with GFP. A double-occupancy PV precedes the formation of individualized L. major PVs, between time points 10:40 and 12:31. Fission is completed after the incorporation of phagolysosomal-membrane in the interface between dividing parasites (time point 12:31). Image acquisition started after 3 h of infection, and the time of acquisition is shown as dhh:mm.