Extended Data Figure 7 ∣. Localization of V5 tagged PfGARP.

a) 3D7-PfGARP KD parasites, in which PfGARP is tagged with the V5 epitope, were grown in the presence of ATc to induce the expression of PfGARP and probed with rabbit anti-glycophorin A (green) and anti-V5 mouse antibodies (red) and counterstained with 4′,6′-diamidino-2-phenylindole (DAPI) to label parasite nuclei. PfGARP tagged with the V5 epitope co-localizes with glycophorin A to the exofacial surface of trophozoite-infected, non-permeablized, RBCs. Scale bar is 5 μm. Data are representative of 3 biologically independent experiments. b) 3D7-PfGARP KD parasites were grown in the presence of ATc to induce the expression of PfGARP. Trophozoite-stage 3D7-PfGARP KD were fixed and incubated with anti-V5 mouse IgG (b) or buffer (c) and probed with anti-mouse IgG labeled with 10 nm gold particles. V5 tagged PfGARP localized to the outer leaflet of 3D7-PfGARP KD infected, non-permeabilized RBCs. Figures labeled with primes represent higher magnifications views of the parent figure. Panels b and c are representative of 2 biologically independent experiments. e) 3D7-PfGARP KD parasites were grown in the presence of ATc to induce the expression of PfGARP and probed with anti-PfCRT (green), anti-V5 antibodies (red) and counterstained with 4′,6′-diamidino-2-phenylindole (DAPI) to label parasite nuclei. PfGARP does not co-localize with PfCRT to the food vacuole in the majority of trophozoite infected RBCs. Panel d is representative of 5 biologically independent experiments.