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CM_21075_sm_SuppFig1.tif1653KFig. S1. Western blotting patterns that confirm the production of triple mutants. (A) ida5oda2 axonemes with and without the tpg1 mutation were analyzed with antibodies against polyglutamylated tubulin (polyE), tyrosinated tubulin (TUB-1A2), TTLL9, and dynein e. (B) Left, ida4oda2 axonemes with and without the tpg1 mutation were analyzed with antibodies against p28 (light chains of dynein a, c, and d) and IC2 (intermediate chain of outer-arm dynein, previously called IC69). The same axonemes were also examined for TTLL9 and the polyglutamylated tubulin (Fig. 1). Middle, ida5oda2 axonemes with and without the tpg1 mutation were analyzed with antibodies against actin and IC2. The same axonemes were also analyzed for TTLL9, dynein e, and polyglutamylated tubulin in A. Right, ida4ida5 axonemes with and without the tpg1 mutation were examined with antibodies against actin (a subunit of inner-arm dyneins, missing in ida5) and p28. The same axonemes were also examined for TTLL9 and polyglutamylated tubulin (Fig. 1).
CM_21075_sm_SuppFig2A.tif2238KFig. S2. Flagellar waveforms in various mutants. High-speed video images taken at 600 frames/sec. The frames are aligned from upper left to lower right, with an interval of 1.67 msec. Bar, 5µm. (A) Wild type and tpg1 cells. (B) ida4 and ida4tpg1. The arrow and arrowhead in the 5th image of wild type denote the principal bend and the reverse bend, respectively. In this example, the ida4tpg1 cell is slowly propagating a shallow principal bend and a large reverse bend. Periods of bend propagation usually lasted only for a few seconds. (C) ida5 and ida5tpg1. The ida5tpg1 cell in this example is propagating a fairly large principal bend and an almost normal reverse bend. Periods of bend propagation lasted for several tens of seconds. The image quality of ida5tpg1 is low because the images were taken with a red filter to avoid photo-induced flagellar adhesion to the glass surface.
CM_21075_sm_SuppFig2B.tif2444KFig. S2. Flagellar waveforms in various mutants. High-speed video images taken at 600 frames/sec. The frames are aligned from upper left to lower right, with an interval of 1.67 msec. Bar, 5µm. (A) Wild type and tpg1 cells. (B) ida4 and ida4tpg1. The arrow and arrowhead in the 5th image of wild type denote the principal bend and the reverse bend, respectively. In this example, the ida4tpg1 cell is slowly propagating a shallow principal bend and a large reverse bend. Periods of bend propagation usually lasted only for a few seconds. (C) ida5 and ida5tpg1. The ida5tpg1 cell in this example is propagating a fairly large principal bend and an almost normal reverse bend. Periods of bend propagation lasted for several tens of seconds. The image quality of ida5tpg1 is low because the images were taken with a red filter to avoid photo-induced flagellar adhesion to the glass surface.
CM_21075_sm_SuppFig2C.tif4299KFig. S2. Flagellar waveforms in various mutants. High-speed video images taken at 600 frames/sec. The frames are aligned from upper left to lower right, with an interval of 1.67 msec. Bar, 5µm. (A) Wild type and tpg1 cells. (B) ida4 and ida4tpg1. The arrow and arrowhead in the 5th image of wild type denote the principal bend and the reverse bend, respectively. In this example, the ida4tpg1 cell is slowly propagating a shallow principal bend and a large reverse bend. Periods of bend propagation usually lasted only for a few seconds. (C) ida5 and ida5tpg1. The ida5tpg1 cell in this example is propagating a fairly large principal bend and an almost normal reverse bend. Periods of bend propagation lasted for several tens of seconds. The image quality of ida5tpg1 is low because the images were taken with a red filter to avoid photo-induced flagellar adhesion to the glass surface.
CM_21075_sm_SuppFig3.pdf40KFig. S3. Amino acid sequence of the microtubule-binding domains (MTBDs) of various heavy chains. Acidic and basic residues are shown in red and blue, respectively. Residues conserved in all dynein heavy chains are shown with a gray background. Cyto1B: Chlamydomonas cytoplasmic DHC. Cyto (Dicty): cytoplasmic dynein of Dictyostelium.

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