Figure 3.

ESCRT-III proteins co-localize during phagocytosis of E. histolytica. (A–D) Representative confocal images of trophozoites in basal conditions and after different times of erythrophagocytosis. PFA-fixed trophozoites were incubated with FITC or TRITC labeled secondary antibodies after incubation with α-rEhVps32 (green) and α-rEhVps2 (red) primary antibodies, respectively; Alexa647 (cyan) and Pacific Blue (blue) directly labeled the α-rEhVps20 and α-rEhVps24 antibodies, respectively. (A) Basal conditions. At right: magnification of white square. (B) Trophozoites at 10 min of erythrophagocytosis. At right: magnification of white square. Pcup: phagocytic cup, e: erythrocytes, red arrow: vesicle stained only by α-rEhVps24 antibody. (C) Confocal representative image of a trophozoite after 90 min of phagocytosis. The areas are marked in squares are magnified in (D): upper panels: erythrocytes covered by EhVps20 and surrounded by EhVps32 and EhVps32 covering two erythrocytes (e) (arrowhead in C) in contrast to others that are surrounded by the protein. Lower panel left: EhVPs32 forming the honeycomb like panel around the erythrocytes together with EhVps24. Lower panel right: EhVps2 and EhVps24 together surrounding the partially digested erythroctyes. phc, phase contrast; sc1, subcomplex 1; sc2, subcomplex 2. Scale bar: 5 μm.