JCS022772 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Analysis of the steady-state level of Cdc16p-HA in SIN mutants. Protein extracts were prepared from the indicated strains, after mid-exponential cultures had been shifted to 36°C for 5 hours before harvesting. (A) Western blot with 12CA5. (B) quantitation of the Cdc16p-HA signal relative to tubulin. (C) Expression of cdc16-HA from the nmt1 promoter, integrated at the leu1 locus, was induced for 20 hours at 25°C. Cells were fixed and stained with DAPI and Calcofluor. (D) Protein samples were prepared from the cells described in C and analysed by western blotting with 12CA5 and antibody to tubulin.

• Supplemental Figure S2 -

  Fig. S2. (A) Characterisation of the antibody against Byr4p. Protein extracts were prepared from the indicated strains and a western blot was probed with affinity-purified antibody against Byr4p or tubulin. (B) Expression of the indicated fragments of byr4 was induced from the nmt1 promoter for 20 hours at 25°C in EMM2 medium in cells expressing Spg1p-GFP. Protein extracts were prepared and analysed by western blotting with antibody against GFP or tubulin. (C) Expression of full-length cdc16-HA integrated at the leu1 locus was induced from the nmt1 promoter for 20 hours at 25°C. Protein extracts were prepared and analysed by western blotting with 12CA5 or antibody against tubulin or Byr4p.