FIG 2.

AmVs accumulate and retain the early endosomal compartment marker Rab5. Indirect immunofluorescence localization of Rab5 using anti-Rab5 antibodies (red) in D. andersoni cells that were synchronously infected with A. marginale labeled with anti-Msp2 antibodies (green) or mock infected with medium (uninfected). Recruitment of Rab5 is not apparent at 1 h postinoculation. Rab5 accumulates and is detected as circumferential staining (arrowhead) on the majority of AmV membranes 5 days postinoculation and is retained at 10 days postinoculation (arrowhead). Inhibition of bacterial protein synthesis (Tetracycline Rx) results in the loss of Rab5 labeling of the AmVs. The uninfected control shows the pattern of staining of Rab5 in D. andersoni cells. The box in the Merge+4′,6-diamidino-2-phenylindole (DAPI) image outlines the area of magnification in the Enlarged column. Scale bars represent 10 μm on a 63×/1.20 objective. The white arrowhead in the first column indicates the location of the AmV and the area enlarged in the inset. All images are at the same magnification, with the exception of the insets and last column, which are enlarged 3-fold.