Figure 5

Immunofluorescence and corresponding brightfield microscopy showing pBS- ap65-neo transfected T. foetus (Tf-pBS- ap65-neo ) expressing AP65 on the surface of non-permeabilized trichomonads (A2) and in hydrogenosomes in permeabilized parasites (B2). The distinct patterns were obtained with the mAbs 12G4 for surface fluorescence and F11 for hydrogenosome fluorescence shown recently to detect the protein in respective compartments (Garcia et al., 2003). T. vaginalis fluorescence patterns (Tv; A4 and B4) were positive controls, and results are identical to those obtained recently (Garcia et al., 2003). No evidence of fluorescence was seen for wild type T. foetus (Tf) and for T. foetus transfected with control plasmid (Tf-pBS-neo). Control hybridoma supernatant lacking anti-AP65 mAb was unreactive with trichomonads.