Fig. 5. Early association of T. cruzi with early endosomes is minimal and precedes lamp-1 acquisition. Immunofluorescence staining of extracellular T. cruzi (arrowheads) following infection of CHO cells transiently expressing (A-C) Rab5-GFP or (D,E) 2xFYVE-GFP for 7.5 minutes. Host cell and parasite DNA is visualized with DAPI (blue). (F) Immunostaining of endogenous EEA1 in T. cruzi-infected CHO cells (15 minutes post-infection). (G) T. cruzi trypomastigotes or (H) fibronectin-coated 3 µm latex beads, incubated with L6E9 myoblasts for 10 minutes were washed extensively to remove uninternalized beads or parasites and further incubated for a total of 60 minutes. Three samples from each condition were fixed at 10, 20, 30 45 and 60 minutes post-incubation and immunostained for EEA1 or lamp-1. Data are represented as means ± s.d.; n=3. Scale bars: 3 µm.