Figure 3. HSP101 delineates a tubular subcompartment of the parasitophorous vacuole.

(a) Live co-localization of HSP101-mCherry (centre left) with a marker protein of the parasitophorous vacuole (GFP^PV, centre). The line in the merge (centre right) indicates profiling of the fluorescent signal (right). Shown are three representative trophozoites demonstrating vacuolar tubules, loops, and vesicles. The blue arrowhead denotes a detached, vacuole-derived, and HSP101-mCherry negative lumen. The red arrowheads denote budding structures at the site of a vacuolar tubule. Note that HSP101-mCherry is excluded from these compartments. (b) FRAP analysis reveals free diffusion from the parasitophorous vacuole to the tubular extensions. Erythrocytes infected with mCherry^PV parasites were analysed by confocal microscopy before (pre bleach) and after (post bleach) photo bleaching (red area, bleach location). Shown is a representative trophozoite and the respective temporal fluorescence analysis in the erythrocyte cytoplasm (blue dotted line); black arrowhead indicates time of the bleaching pulse. Scale bar, 5 μm.