PhIL1-interacting protein, PIP1, is located at the gametocyte IMC and is essential for gametocyte development. (A) PIP1-HA-glmS transfectants were labeled with anti-PhIL1 rabbit (green) and anti-HA (red) at different developmental stages (II±V). Nuclei were labeled with DAPI. There is an apparent reduction in fluorescence intensity of PIP1-HA labeling at the tips of the gametocyte in stage IV (yellow arrow). In stage V, the fluorescence becomes punctate at the parasite periphery, while PhIL1 remains homogenous. (B) 3D-SIM imaging of PIP1-HA-glmS during gametocyte development. Homogeneous labeling of the IMC plates by anti-HA (red) are observed (yellow arrows) in the early stages of development. During stage III±IV of development, there is a loss of labeling from the plates near the tips of the elongated parasite (blue arrows), with some protein labeling of the tips themselves (white arrows). Scale bar: 5 μm.

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