FIG 3.

AmVs accumulate and retain the recycling endosomal compartment marker Rab4. Indirect immunofluorescence localization of Rab4 using anti-Rab4 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 Rab4 is not apparent 1 h postinoculation. Rab4 accumulates on the majority of AmVs 5 days postinoculation and is retained at 10 days postinoculation. Inhibition of bacterial protein synthesis (Tetracycline Rx) results in loss of Rab4 labeling of the AmVs. The uninfected control shows the pattern of staining of Rab4 in D. andersoni cells. The box in the Merge+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.