JCS03352 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

  Adobe PDF

• Supplemental Table S2 -

  Adobe PDF

• Supplemental Figure 1 -

  Fig. S1. Additional motility traces of TbCMF mutants. Additional mutants from each class were analyzed via motility traces as described in the text. n=50-60.

• Supplemental Figure 2 -

  Fig. S2. GFP alone does not localize specifically to the flagellum. Fluorescence microscopy of live cells (top) and detergent-extracted cytoskeletons (bottom) from cells expressing GFP alone. When expressed from pKH10 (left), the same vector used for fusions (Fig. 6), fluorescence is evident in the cell body but not in the flagellum of live cells and is lost upon detergent extraction. The pKH10 vector was used specifically to give low-level expression. When GFP alone is expressed from pLEW82 (right), which employs a strong T7 promoter (Wirtz et al., 1998) GFP is primarily in the cell body, with minor fluorescence in the flagellum and this is lost upon detergent extraction (bottom).

• Movie 1
• Movie 2
• Movie 3
• Movie 4
• Movie 5 -

  Movies 1-5. Loss of TbCMF genes has varied effects on flagellar motility. Ten-second real-time movie captures for TbCMF mutants from class 2 (TbCMF 3, Movie 1), class 3 (TbCMF 46, Movie 2) and class 4 (TbCMF 5, Movie 4; and TbCMF 9, Movie 3). Mutants display motility irregularities corresponding to the severity of the clustering phenotype seen in whole culture (see text for details). Movie 5 shows uninduced TbCMF 5 cells and is provided as an example of WT flagellar beat.