PhIL1 is located at the IMC in gametocytes. (A) Immunofluorescence microscopy showing anti-PhIL1 (green) at the periphery of a 3D7 stage IV gametocyte, showing fluorescence close to the anti-β-tubulin (red) labeling. Immunofluorescence microscopy of stage IV gametocytes revealed a strong PhIL1 signal at the periphery, where it is closely associated with β-tubulin and was located close to the known IMC marker, the glideosome-associated protein 50 (B) Immunofluorescence microscopy of a stage IV gametocyte showing overlap of GAP50-GFP (red) and PhIL1 (green) at the periphery of the cell. (D, E) Immunofluorescence microscopy of stage IV PhIL1-GFP (D) and PhIL1-HA (E) gametocyte transfectants. Parasites were labeled with anti-PhIL1 (green) and anti-GFP or anti-HA (red). Nuclei were labeled with DAPI. Scale bars: 5 μm. A PhIL1-HA line was also generated by integrating 3xHA into the 3' end of the PhIL1 locus. Immunofluorescence signals for HA and GFP overlapped with the signal from anti-PhIL1 antibody, confirming the correct location of the tagged proteins (D and E).

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