FIG 4.

Expression and localization of nonphosphorylatable (S147A) and phosphomimetic (S147D) EhCoactosin variants. HM-1:IMSS strain trophozoites were transfected with expression vectors designed for tetracycline-regulated EhCoactosin expression with an N-terminal hemagglutinin (HA) epitope tag fused to either wild type (Coac-WT), S147A (Coac-A), or S147D (Coac-D). (A) Western blot. Protein expression was induced with tetracycline followed by immunoprecipitation (IP) and blotting with an anti-HA antibody. The expected protein size was 17 kDa. (B) Localization of EhCoactosin and S147A and S147D variants during erythrophagocytosis. Trophozoites expressing each EhCoactosin variant protein were incubated with human erythrocytes and then prepared for fluorescence microscopy by staining F-actin with phalloidin (green) and the EhCoactosin proteins with a mouse anti-HA monoclonal antibody. The Coac-WT, Coac-A, and Coac-D proteins all localized to the phagocytic cup. Bars, 10 μm.