(A) Transfectants (ITK strain) expressing the PfEMP1B-GFP chimera were co-labeled with BODIPY-ceramide and imaged using 3D-SIM. Higher magnification images are displayed in the right column. PfEMP1B-GFP is present in horseshoe-shaped Maurer’s clefts (aqua arrows). Spherical structures with no GFP labeling are marked with yellow arrows. (B) EqtII-permeabilized PfEMP1B-GFP transfectants were probed with antibodies recognizing GFP and REX1 and imaged by 3D-SIM. (C-F) EqtII-permeabilized 3D7 strain or PfEMP1B-GFP transfectants were labeled with anti-PfEMP1 ATS (C) or anti-GFP (D-F) and 6 nm Protein A-gold, and prepared for EM. A tether (t) and the parasite (P) are indicated. Scale bars = 2 μm (A, first three columns, B), 1 μm (A, last column), 500 nm (B, last column), 100 nm (C-F).