FIG. 2.

Epitope-tagged GRA14 traffics to the PV and localizes to the IVN and PVM. (A) Diagram of the GRA14-HA construct showing that expression is driven from the endogenous promoter and indicating the position of the HA epitope tag at the extreme C terminus of the protein. A heterologous GRA2 3′ untranslated region was used, and the selectable marker HPT is not shown. (B) IFA of infected cells (with a clonal parasite line stably expressing GRA14-HA) demonstrating that GRA14-HA traffics correctly to the PV and colocalizes with GRA7. (C and D) Immunoelectron microscopy was used to examine cells infected by GRA14-HA parasites. P, parasite; DG, dense granule; HC, host cell. (C) GRA14-HA labeling was found inside the cores of the dense granules. Two different parasite images are shown (scale bars, 100 nm in the first image and 200 nm in the second image). (D) GRA14-HA labeling was also found on the IVN (single arrowhead) and on the PVM (double arrowhead) (scale bar, 200 nm).