PIP2 is absent from malaria-induced vacuoles. (A,B) Fluorescence micrographs of a GFP-PLCδ1 PH domain–loaded ghost (green) infected by a young ring-stage malaria parasite. The parasite nucleus is stained with blue Hoechst 33342 as shown in the merged image. Scale bar equals 2 μm. (C,D). Erythrocyte ghost loaded with Alexa-Fluor 594–labeled annexin V (red) containing a Hoechst-stained intracellular ring-stage parasite as shown in the merged image. Scale bar equals 2 μm. (E,F). Merozoite (blue) attached to the surface (as shown by light microscopy) (E) and indenting (arrowhead) the membrane of an erythrocyte ghost loaded with GFP-PLCδ1 PH domain (F). Scale bar equals 2 μm. (G-H). An erythrocyte ghost loaded with GFP-PLCδ1 PH domain (G) containing a recently invaded young ring-stage parasite (H) (merge with Hoechst stain). Images acquired with a 100×/1.35 oil objective (Olympus).
