Figure 5.
The PKD2–mastigoneme complexes associate with DMT4 and DMT8. (A and B) Tomographic slices of an intact Chlamydomonas wild-type cilium viewed in longitudinal section (A, thickness = 25 nm) and cross section (B, thickness = 5 nm). Mastigonemes (red arrowheads) are positioned closest to DMT4 and DMT8. The white lines labeled B in A indicate the position of the cross section shown in B. The cross section (B) shows the relative positions of the two rows of mastigonemes to the ciliary beating direction (small white arrows). For clarity, DMTs 4 and 8 are highlighted by cloning the subtomogram average of the 96-nm DMT repeat back into the raw tomogram for all axonemal repeats along DMT4 and 8. Bar = 100 nm. (C) Tomographic slices of the averaged mastigoneme base (left, thickness = 2 nm; bar = 10 nm) and after applying threefold symmetry (right, thickness = 2 nm; bar = 5 nm) from intact Chlamydomonas cilia. The averaged structure is based on 53 particles extracted from six tomograms. The ciliary membrane (M) and glycocalyx (G) are indicated. The red arrow indicates variable positions of the mastigonemes with respect to the base in our samples. (D) Tomographic slice (left, thickness = 2 nm; bar = 5 nm) and 3D isosurface rendering representation (right) of the averaged mastigoneme base from demembranated cilia (axonemes) after applying threefold symmetry. The dashed lines on the left indicate the levels of the membrane (M) and glycocalyx (G); the dashed lines on the right mark the trans-membrane (M), collar, and neck (N) subregions of the mastigoneme base and the attached mastigoneme filament (F). The dimensions of the averaged base complex are shown in Fig. S5 F. (E) Tomographic slices (thickness = 5 nm; bar = 50 nm) of demembranated cilia showing thin linkers (arrowheads) between the mastigoneme bases and DMTs; the DMT number is indicated. See Fig. S5, C–E for further details of contact site between the base complex and the DMTs. (F and G) 3D isosurface rendering of a representative region of an intact Chlamydomonas wild-type cilium (same as shown in A and B). Subtomogram averages of the mastigoneme filaments, 96-nm axonemal DMT repeats, and the mastigoneme base complexes were cloned back into the positions of individual repeat units in the raw reconstruction; selected substructures were segmented and colored as indicated for better visualization.

The PKD2–mastigoneme complexes associate with DMT4 and DMT8. (A and B) Tomographic slices of an intact Chlamydomonas wild-type cilium viewed in longitudinal section (A, thickness = 25 nm) and cross section (B, thickness = 5 nm). Mastigonemes (red arrowheads) are positioned closest to DMT4 and DMT8. The white lines labeled B in A indicate the position of the cross section shown in B. The cross section (B) shows the relative positions of the two rows of mastigonemes to the ciliary beating direction (small white arrows). For clarity, DMTs 4 and 8 are highlighted by cloning the subtomogram average of the 96-nm DMT repeat back into the raw tomogram for all axonemal repeats along DMT4 and 8. Bar = 100 nm. (C) Tomographic slices of the averaged mastigoneme base (left, thickness = 2 nm; bar = 10 nm) and after applying threefold symmetry (right, thickness = 2 nm; bar = 5 nm) from intact Chlamydomonas cilia. The averaged structure is based on 53 particles extracted from six tomograms. The ciliary membrane (M) and glycocalyx (G) are indicated. The red arrow indicates variable positions of the mastigonemes with respect to the base in our samples. (D) Tomographic slice (left, thickness = 2 nm; bar = 5 nm) and 3D isosurface rendering representation (right) of the averaged mastigoneme base from demembranated cilia (axonemes) after applying threefold symmetry. The dashed lines on the left indicate the levels of the membrane (M) and glycocalyx (G); the dashed lines on the right mark the trans-membrane (M), collar, and neck (N) subregions of the mastigoneme base and the attached mastigoneme filament (F). The dimensions of the averaged base complex are shown in Fig. S5 F. (E) Tomographic slices (thickness = 5 nm; bar = 50 nm) of demembranated cilia showing thin linkers (arrowheads) between the mastigoneme bases and DMTs; the DMT number is indicated. See Fig. S5, C–E for further details of contact site between the base complex and the DMTs. (F and G) 3D isosurface rendering of a representative region of an intact Chlamydomonas wild-type cilium (same as shown in A and B). Subtomogram averages of the mastigoneme filaments, 96-nm axonemal DMT repeats, and the mastigoneme base complexes were cloned back into the positions of individual repeat units in the raw reconstruction; selected substructures were segmented and colored as indicated for better visualization.

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