JCS03464 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Measurement of individual bipolar cells during an HU block. Briefly, cdc25-22 cells were blocked for 2 hours at the restrictive temperature and then released to the permissive temperature in the presence of 12 mM HU. Cells were stained for actin every 30 minutes and the length of bipolar cells was measured. 82 cells were measured.

• Supplemental Figure S2 -

  Fig. S2. Branch position depends on the nucleus in the absence of microtubules. (A) Correlation between number of nuclei and number of branches per cell in cdc11-119 cells treated with MBC after 0, 2 and 4 hours block. (B) FACS profile and images of Cdc13 switch-off cells treated with MBC. (C) Quantification of branch position in cdc11-119 cells after centrifugation. MBC was added at time of centrifugation. In the graph, 0 is the cell centre and 1 the cell tip. (D) DAPI staining of cdc11-119 cells treated with MBC for 3 hours after centrifugation.

• Supplemental Figure S3 -

  Fig. S3. Cdc25-22 cells blocked for 3 hours were stained for DAPI (top), and cdc25-22 cells blocked for 3 hours were stained for microtubules (bottom).

• Supplemental Figure S4 -

  Fig. S4. Effect of TBZ and MBC on cell polarity. Cdc10-129 cells were blocked for 3 hours at 36.5°C prior to drug treatment (t=0). Cells were treated with DMSO, 50 μg/ml MBC, 100 μg/ml TBZ and 50 μg/ml MBC plus an 8-minute pulse of 12.5 mM latA. Treatment with MBC causes very inefficient branching. A pulse of latA in the presence of MBC induces cell branching at similar levels to TBZ treatment.