Colocalization studies with anti-peptide 1 antibodies and fixed 3D7 pRBC from schizont ghosts and PfEMP3 truncated 3D7 cell lines. (A) Schizont ghosts. (B) PfEMP3 truncated 3D7 cell lines. Parasite nuclei were stained with DAPI (blue). Double staining was carried out with anti-peptide 1 rabbit serum (green) and mouse serum (B28) against the 77-kDa carboxy terminus of MC protein PfSBP1 (red) or mouse serum (DG662) against the carboxy terminus of PfEMP3 (red). Anti-peptide 1 serum was detected by using FITC-conjugated goat anti-rabbit IgG. Colocalizing antibody was detected by using Texas red-conjugated goat anti-mouse IgG. Magnification, x1,000. A truncated form of PfEMP-3 obtained by transfection was no longer distributed around the cytoplasmic surface of the erythrocyte membrane but was instead associated with structures believed to be MC. IFA experiments with antibodies against PfEMP-3 and STEVOR showed the colocalization of truncated PfEMP-3 and STEVOR (B), further supporting the observation that STEVOR is located in MC.

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