Expression of PfEMP1-GFP chimeras inP falciparum–infected RBCs. (A) Fluorescence images of 3D7-TmATS at different stages. The first row represents the GFP fluorescence; second row, the DAPI (4,6 diamidino-2-phenylindole)–stained nuclei; third row, an overlay of the GFP and DAPI images; fourth row, the bright-field images; last row, overlays of all 3 images. (B) ER localization of GFP fluorescence in 3D7-TmATS transfectants. First panel, GFP fluorescence; second panel, ER tracker fluorescence; third panel, overlay of GFP and ER tracker fluorescence; fourth panel, bright-field images. (C) Immunofluorescence imaging of fixed 3D7-ACPTmATSs transfectants. First row, αGFP; second row, αEXP1; third row, bright-field image; fourth row, overlay of all images. (D-E) Expression of K119TmATS-GFP, K60TmATSs-GFP, and K60Tm(ama1)ATSs-GFP proteins. All images represent live parasites of the 3D7 or D10 strains collected at ring, trophozoite (Troph), and schizont stages during the 48-hour asexual life cycle and were stained with DAPI. First row, GFP fluorescence; second row, DAPI; third row, bright-field images; fourth row, overlay of GFP fluorescence and bright-field image. DNA in early ring stages did not stain with DAPI; only nuclei of parasites more than 12 hours past invasion were visible after DAPI staining.