FIG 4 .

The phospholipid composition of the HCM appears to be altered during liver stage egress. HeLa cells were transfected with constructs encoding the PH domain of phospholipase C-δ1 fused to GFP to visualize PIP₂ (A), the PH domain of AKT serine/threonine kinase 1 fused to GFP to visualize PIP₃ (B), the lactadherin-C2 domain fused to GFP to visualize PS (C), and an ezrin-GFP fusion protein (D). Transfected cells were subsequently infected with parasites expressing mCherry fused to the PVM marker protein ExpI (red), and parasite development was monitored by live-cell time-lapse microscopy. The localization of the phospholipid reporters and ezrin (green) in infected cells with intact PVM and in the same cells with a ruptured PVM is shown. Results are representative of 26 (PIP₂), 15 (PIP₃), 19 (PS), and 20 (ezrin) analyzed egress events in two or three independent imaging experiments. All scale bars, 10 µm. See also Movies S4 and S5.