Co-localization and association of EhNPC1 and EhNPC2 with cholesterol and endosomal molecules.

<p>Trophozoites were incubated with erythrocytes as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006089#ppat.1006089.g007" target="_blank">Fig 7</a> and processed for confocal microscopy after incubation with α-EhNPC1 and α-EhNPC2 antibodies and (A) Lysotracker or (B) α-LBPA or (C) α-EhADH antibody and corresponding secondary antibodies. (a-d,g,h) Magnification of white squares. (e,f,i,j) Magnifications from other images. Ph c: phase contrast images, arrow in I: EhNPC1 outside endosomes, arrowhead: EhNPC2 inside endosomes. D, E) Trophozoites in basal condition or after 5+60 min of erythrophagocytosis were lysed and immunoprecipitation assays (IP) were performed using α-EhNPC1 (D) or α-EhNPC2 (E) antibodies or preimmune serum (PS). Immunoprecipitated proteins of trophozoites were analyzed by western blot and dot blot experiments, using α-EhNPC1, α-EhNPC2, α-EhADH, α-EhRab7A, α-EhSERCA and α-cholesterol antibodies.