Eps15 is located at sites containing host cell cytosol filled vesicles and cytostomal neck structures. (A-C) Three cells expressing endogenously tagged Eps15-2xFKBP-GFP analysed using CLEM, showing confocal images and EM sections of the same cell (A, B, EM slices of the entire cells were captured, C, only partial capture of slices). (A) Additional information on the cell shown in Fig. 3. Left: maximum intensity projection of a 3D reconstruction of the Eps15-GFP signal overlayed over DIC (confocal microscopy). xy and yz projections show position of the Eps15 foci in the cell and position of the EM slices shown to the right. Foci are specified by arrow color. Black arrows: cytostomal neck-like structures found in the top planes of the cell, suggesting the host cell cytosol filled structures indicated by light blue arrows might be cytostomes. (B) Left: maximum intensity projection of a 3D reconstruction of the Eps15-GFP signal overlayed over a single DIC slice (confocal microscopy). xy and yz projections show position of the Eps15 foci in the cell. Black arrows: cytostomal neck-like structures in bottom planes of the cell, suggesting the host cell cytosol filled structures indicated by light red arrows might be cytostomes. (C) EM sections recovered from the infected RBC show to the left (DIC/GFP overlay of a single z-slice obtained by confocal microscopy) with 2 Eps15-GFP foci. Red arrow: focus corresponding to the region with the host cell cytosol-filled structures. Black arrows: cytostomal neck structures connected to the vesicles. FV, food vacuole; V, vesicle containing material with similar density to host cell cytosol. Size bars confocal images: 2 µm, EM images: 0.5 µm and 0.25 µm in the EM enlargements. A Gaussfilter was used for the green signal in (A)
and (B).
Birnbaum J, Scharf S, Schmidt S, Jonscher E, Hoeijmakers WAM, Flemming S, Toenhake CG, Schmitt M, Sabitzki R, Bergmann B, Fröhlke U, Mesén-Ramírez P, Blancke Soares A, Herrmann H, Bártfai R, Spielmann T. A Kelch13-defined endocytosis pathway mediates artemisinin resistance in malaria parasites. Science. 2020 Jan 3;367(6473):51-59. PMID: 31896710