JCS03078 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Semi-quantitative RT-PCR performed on RNA samples extracted using the Trizol method from the indicated cell lines. For each experiments, two pairs of primers were used, one for amplification of the target RNA and the other for the aldolase mRNA that serves as internal control. Position of each amplification product is shown by arrows.

• Supplemental Figure 2 -

  Fig. S2. Cross-sections through the flagella of wild-type, non-induced (A) or induced (TbPF20)RNAi mutants (B). These images show no particular differences in the amount of cell body to flagellum interaction in all three conditions.

• Supplemental Figure 3 -

  Fig. S3. Endocytosis is not modified in cells from unshaken cultures of control, (TbPF16)RNAi or snl2 cells. Top, labelling of the endosomal compartment by RAB5. Bottom, labelling of the recycling compartment by RAB11. The flagellar pocket is found at the posterior end of the cell and appears functional in all cases.

• Movie 1 -

  Movie 1. Non-induced (TbDNAI1)RNAi mutant. The cell on the right swims forward with its flagellum leading, showing exclusively tip-to-base wave propagation. The cell on the left is seen changing orientation, with base-to-tip wave propagation.

• Movie 2 -

  Movie 2. (TbPF16)RNAi mutant induced for 2 days. The first cell has an attached flagellum and displays very little flagellum movement. The second cell has two flagella, the new one is paralysed and detached from the cell body (red) and the old one (tip pointing towards the left) is still attached and motile.

• Movie 3 -

  Movie 3. (TbDNAI1)RNAi mutant induced for 2 days. This cell only displays base-to-tip flagellar wave propagation.

• Movie 4 -

  Movie 4. (TbPFR2)RNAi mutant induced for 2 days. Four different cells are presented. Movement is still observed at the distal tip of the flagellum but not transmitted to the cell body. The posterior end looks straight and longer compared to non-induced controls. Waves seem to constantly alternate between base-to-tip and tip-to-base (bottom right cell).

• Movie 5 -

  Movie 5. Non-induced (TbPF16)RNAi mutant. At the end of cytokinesis, daughter cells are only joined by their posterior end and their motile flagella point in opposite direction as the whole edifice is twisted. No net movement of the complex is visible.

• Movie 6 -

  Movie 6. (TbPF16)RNAi mutant induced for 2 days. At the same stage of cytokinesis, only the old flagellum is still motile (left). Its beating leads to motility, what drags the cell with the immotile new flagellum.

• Movie 7 -

  Movie 7. (TbPF16)RNAi mutant induced for 3 days. Both flagella are non-motile and progression through cytokinesis is significantly slowed down.

• Movie 8 -

  Movie 8. snl-2 mutant induced for 2 days. Both flagella show reduced motility (the old one is probably on the left, the new one on the right) but some movements still occur and the edifice can twist to a certain degree.