JCS035626 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Basal body replication takes place in the absence of flagellum in 48-hour-induced IFT88^RNAi cells. Merged images of double IFA staining with mAb22 (marker of the proximal end of the mature and the pro-basal body, green) and the anti-TBBC antibody (marker of the mature basal body, red) of detergent extracted cytoskeletons from two nonflagellated cells stained with DAPI (blue). Inset shows magnification of the basal body region.

• Supplemental Figure S2 -

  Fig. S2. The FPC is disorientated as soon as cells fail to assemble a normal flagellum. (A-B) Cold Triton X-100 treatment of a control cell (A) or IFT140^RNAi cells induced for 72 hours (B). In controls, the two flagella emerge from separate FPCs that are parallel. In cells that possess an old flagellum and an aberrantly short new flagellum, the orientation of the FPC associated to the new flagellum is not fixed relative to the long axis of the cell or to the orientation of the FPC of the old flagellum.

• Supplemental Figure S3 -

  Fig. S3. BILBO1 organisation is modified in non-flagellated cells. IFT88^RNAi cells were noninduced (−TET) or induced for 3 days (+TET) and stained by IFA with the mAb25 monoclonal antibody recognising an axonemal protein (red) and with an antibody recognising BILBO1 (green). Samples were counterstained with DAPI (blue). BILBO1 staining is less well defined in nonflagellated cells.

• Supplemental Figure S4 -

  Fig. S4. Organisation of the lysosomes, endoplasmic reticulum and glycosomes in cells treated with IFT siRNA. Cells from the indicated cell lines were non-induced (−TET; A,C,E) or induced for 3 days (+TET; B,D,F) and stained by IFA with antibodies against p67 (lysosome marker, red, A,B), against BiP (endoplasmic reticulum marker, green, C,D) and against aldolase (glycosome marker, yellow, E,F). Samples were counterstained with DAPI (blue).