GFP-expression in blood stages of three reporter <i>P</i>. <i>falciparum</i> parasite lines (GFP@<i>cam</i>, GFP@<i>gapdh</i>, GFP@<i>hsp70</i>).

<p>A. Fluorescence microscopy of different blood stages. R: rings; T: trophozoites; ES: early schizonts; LS: late schizonts; G: gametocytes. Nuclei were stained with the DNA-specific dye Hoechst 33342. All pictures were recorded with standardized exposure/gain times to visualize differences in fluorescence intensity (GFP 0.7 s; Hoechst 0.136 s; bright field 0.62 s (1x gain)). In <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168362#pone.0168362.s003" target="_blank">S3</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168362#pone.0168362.s005" target="_blank">S5</a> Figs. the complete set of microscope images are shown. B. Fluorescence intensity of rings (16 h), (late) trophozoites (30 h) and schizonts (42 h) as determined by flow cytometry. Infected red blood cells (RBC) were stained with the DNA-specific dye Hoechst 33258 to distinguish infected RBC from uninfected RBC and rings (Gate G1), trophozoites (Gate G2) from schizonts (Gate G3). Left side panels show dot plots of both Hoechst fluorescence intensity (DNA content) and GFP fluorescence intensity. Right side panels show GFP fluorescence intensity from parasites with either G1 and G2 (black) gate or G3 (red). MFI: mean fluorescence intensity and the black- (rings and trophozoites) and red- (schizonts) bar show the region selected to calculate the MFI.