Figure 6 | Scientific Reports

Figure 6

From: A Plasmodium plasma membrane reporter reveals membrane dynamics by live-cell microscopy

Figure 6

Double-fluorescent parasites reveal interactions of the PPM with the parasite ER during oocyst and liver stage development. (a) SBFSEM of the P. berghei ER. HeLa cells infected with mCherry-expressing parasites were fixed at 48 hpi and osmium-stained for EM. Cells were vertically cut and images were taken by SBFSEM. The boxed area is shown at a higher magnification on the right side and shows an ER extension (red arrow) to the parasite PM and or PVM (white arrow). P, parasite; asterisks, parasite nuclei. (b) Schematic representation of the pL0017CsfGFP-PbSec61β-CPbPMP1-mCherry plasmid. The sfGFP and mCherry fusion proteins were both expressed under the control of the constitutive eef1α promoter. The 3′-UTR was taken from Pbdhfr/ts. (c) and (d) Interactions of the parasite ER and the PPM in oocysts and liver stage parasites. (c) Midguts of sfGFP-PbSec61β/PbPMP1-mCherry parasite-infected mosquitoes were isolated at day 7 after the infectious blood meal and were analyzed live by confocal microscopy. (d) HeLa cells were infected with sfGFP-PbSec61β/PbPMP1-mCherry parasites and analyzed live by confocal microscopy at 24 hpi (upper row) and 48 hpi (two lower rows). SfGFP-PbSec61β (green), PbPMP1-mCherry (red). Extensions of the ER in contact with the surrounding PPM were found in all oocysts and liver stages examined (a total of 20 oocysts analyzed at day 7 and 9 post-feed, a total of 60 liver stages at 24 hpi and a total of 60 liver stages at 48 hpi assessed). For confocal z-stacks see also Supplementary Movies S4 and S5. For a time-lapse movie of sfGFP-PbSec 61β/PbPMP1-mCherry parasite liver stage development see also Supplementary Movie S6. Scale bars correspond to 10 µm, if not labelled differently.