Co-localization of EhVps32, Gal/GalNac lectin, actin and Lysotracker during erythrophagocytosis.

Trophozoites were incubated with erythrocytes for different times and analyzed through confocal microscopy. (A) After erythrophagocytosis, trophozoites were incubated with rabbit αrEhVps32 and mouse αGal/GalNac lectin antibodies and rhodamine-phalloidin, and then, with α-rabbit FITC and α-mouse Pacific blue-labeled secondary antibodies, respectively. Arrowheads: co-localization at phagocytic cup. Arrows: co-localization at phagosome membrane. e: erythrocytes. (B) Pearson’s coefficient (PC) to quantify EhVps32 and Gal/GalNac lectin co-localization in the entire cell (total) and in the plasma membrane (PM). (C) Confocal microscopy of trophozoites incubated with Lysotracker and treated with αrEhVps32 and FITC-labeled secondary antibodies. Arrows: co-localization of EhVps32 and Lysotracker. Arrowheads: Lysotracker-stained phagosomes without EhVps32 signal. e: erythrocytes. (D) Pearson’s coefficient (PC) to quantify EhVps32 and Lysotracker co-localization in the entire cell. (*) p<0.05 and (**) p<0.01. (E) Proportion of erythrocytes inside phagosomes decorated by αrEhVps32 antibodies and Lysotracker with relation to total ingested erythrocytes per trophozoite.