JCS024224 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Cellular levels of mis4 mRNA were not decreased by TSA treatment. Wild type and mis4-242 mutants were cultured in rich medium at 26°C, and at the indicated time points after addition of 12.5 mg/ml TSA samples were taken to extract RNAs. The levels of mis4 mRNA and ura4 mRNA levels were determined by northern blotting. Ribosomal RNAs stained with ethidium bromide were also shown. The band intensities were quantified by Phosphoimager (BioRad) and the ratio of mis4 mRNA to ura4 mRNA at each times was expressed in arbitrary units mis4/ura4 (a.u.).

• Supplemental Figure S2 -

  Fig. S2. The timecourse of the re-formation of the 20S APC/C complex by TSA treatment in cut4-533 mutants. (A) Wild type and cut4-533 mutants grown exponentially in YPD liquid medium at 26°C. TSA (12.5 µg/ml) was added to the cut4-533 cultures. The cells were harvested at the indicated time points after TSA addition. A 15-40% sucrose gradient centrifugation was carried out and the cell extracts were fractionated into 20 tubes. The second to twelfth fractions were used for western blotting with Cut4 and Cut9 antibodies. Arrowheads indicate the fast migrating bands of Cut4. (B) Signal intensities of the fast migrating bands of Cut4 and the bands of Cut9 in A were measured and plotted in bar graphs.