Fig. 3: Defective axonemal assembly in RBPm1-null male gametogenesis.

A Detection of formation and exflagellation of axonemes during male gametogenesis (0, 8, and 15 mpa) by staining α-Tubulin (left panels) and β-Tubulin (right panels). Nuclei were stained with Hoechst 33342. Four independent experiments with similar results. Scale bars: 5 µm. B Immunoblot of α- and β-Tubulins in gametocytes. The numbers indicate the relative intensities of the bands in the immunoblots. BiP as a loading control. Two independent experiments with similar results. C Ultrastructure expansion microscopy (U-ExM) of the axonemes in male gametocytes stained with α-Tubulin antibody at 8 mpa. Three independent experiments with similar results. Scale bars: 5 μm. D Transmission electron microscopy of axoneme architecture in male gametocytes at 8 mpa. Inset panels show longitudinal sections (top panels) and cross sections (bottom panels) of axonemes. The enclosed area (black box) was zoomed in. Pie charts show the quantification of axoneme (“9 + 2” microtubules) in the mutant parasites. n is the total number of intact and defective axoneme structures observed in each group. Three independent experiments with similar results. Scale bars: 1 μm.