Skip to main content
. 2016 Apr 11;7:11191. doi: 10.1038/ncomms11191

Figure 9. Retromer is required to maintain TgHP03 in the parasite membrane.

Figure 9

(a) Schematic representation of TgHP03 topology inside the parasite membrane. (b) Confocal imaging of extracellular parasites with knock-in TgHP03-cMyc in the iKoTgVps35 mutants. Immunofluorescence assay (IFA) was performed in the absence of ATc and detergent permeabilization. Nuclei of the parasites as stained with 4′, 6-diamidino-2-phenylindole (DAPI; blue). Bar, 2 μm. (c) Magnified image of an extracellular parasite expressing TgHP03-cMyc protein in the iKoTgVps35 mutants grown in the absence of ATc and detergent permeabilization (upper, left panel); intracellular parasites expressing TgHP03-cMyc protein in the iKoTgVps35 genetic background. IFA was performed in the absence of ATc and in the presence detergent permeabilization (upper, right panel); magnified image of an intracellular parasites expressing TgHP03-cMyc protein in the iKoTgVps35 genetic background in the presence of ATc and detergent permeabilization (lower, left panel) and intracellular parasites expressing TgHP03-cMyc protein in the iKoTgVps35 genetic background in the presence of ATc and detergent permeabilization (lower, right panel). Bar, 2 μm. (d) Immunoblots of parasites expressing TgHP03-cMyc protein in the knock-in iKoTgVps35 mutants grown in the presence or absence of ATc. ENO2 was used as a loading control. Molecular weights (kDa) were shown on left. (e) Quantification of TgHP03 levels in these parasites expressing TgHP03 protein in the knock-in iKoTgVps35 mutants that were grown in the absence or presence of ATc. (f) The surface localization of glycosyl–phosphatidyl inositol (GPI)-anchored SAG1 and SAG3 were determined in iKoTgVps35 mutants in the presence or absence of ATc using monoclonal antibodies specific to SAG1 and SAG3. Bar, 2 μm.