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

<p>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. (*) <i>p</i><0.05 and (**) <i>p</i><0.01. (E) Proportion of erythrocytes inside phagosomes decorated by αrEhVps32 antibodies and Lysotracker with relation to total ingested erythrocytes per trophozoite.