Fig. 4

Expression and intracellular localization of dUTPase at different stages of the intraerythrocytic cycle. The intracellular localization of dUTPase was assessed by live cell fluorescence microscopy of cells overexpressing PfdUTPase fused to GFP. a In vivo microscopy of parasites transfected with the plasmid pHH2-Pfdut-GFP. Cells were previously stained with Hoechst33342 and Mitotracker for nucleus and mitochondria localization. Images were taken with a confocal Leica TCS SP5 microscopy system and show a single optical plane. b Western blotting of extracts of parasites overexpressing PfdUTPase-GFP using a polyclonal anti-PfdUTPase antibody. A band of approximately 46.5 kDa appears only in transfected parasites (PfdUTPase GFP OE), corresponding to the fusion protein dUTPase-GFP. c Expression of PfdUTPase in 3D7 wild-type cells throughout the intraerythrocytic cycle. Protein extracts of parasites in ring (R), trophozoite (T) and schizont (S) stages were subjected to Western blotting with the polyclonal dUTPase antibody. Anti-Hsp70 was used as loading control