Fig. 2: APR2 localizes at APR throughout zygote to ookinete development.

a IFA of APR2 expression during zygote (stage I) to ookinete (stage V) development of two HA-tagged strains 6HA::apr2 (left panel) and apr2::6HA (right panel). Three independently performed experiments with similar results. Scale bars: 5 μm. b Fluorescence detection of APR2 in living parasites during zygote to ookinete development of two GFP-tagged strains gfp::apr2 (top panel) and apr2::gfp (bottom panel). Two independently performed experiments with similar results. Scale bars: 5 μm. c Co-localization analysis by IFA for APR2 with proteins of known cellular localizations in ookinetes. P28 (plasma membrane, PM), GAP45 (inner membrane complex, IMC), α- and β-Tubulin (subpellicular microtubule, SPMT), MyosinB and SAS6L (apical tubulin ring, ATR), chitinase and CTRP (microneme). Top panel shows a diagram of apical structure of ookinete. APR2 is tagged with a 6HA. SAS6L and MyosinB are tagged with a 4Myc. P28, GAP45, α- and β-Tubulin, chitinase, and CTRP are detected using the antibody or antiserum. Two independently performed experiments with similar results. Scale bars: 5 μm. d Immuno-EM images of the apr2::6HA ookinete. Gold particles coupled with the anti-HA antibody are located at APR (white dashed lined area) in the longitudinal view (left) and cross view (right). Two independently performed experiments with similar results. Scale bars: 0.5 μm. e Ultrastructure expansion microscopy (U-ExM) of APR2 in the apr2::6HA ookinetes from 3- and 12-h in vitro culture. The parasites were co-stained with the anti-HA antibody and the protein NHS-ester dye. Dashed-lined area containing NHS-ester dense region (triangle) were zoomed in and shown in the right panels. Three independently performed experiments with similar results. Scale bars: 5 μm.