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. 2006 May 1;20(9):1162–1174. doi: 10.1101/gad.1367206

Figure 4.

Figure 4.

G2/M delay in top2-B44 is not enforced by Pds1/Securin. (A) Cell cycle analysis of double and triple mutants of top2∷top2-B44 combined with pds1Δ or pds1Δ mad2Δ. Since the pds1Δ cells are temperature-sensitive slightly above 28°C, the G1 synchrony was performed at 26°C, then upon release from G1 the temperature was shifted up to 28°C. (B) Cell cycle analysis of top2∷top2-B44 pds1Δ in combination with GAL1-ESP1-GFP-NLS (Jensen et al. 2001). Cells were synchronized in medium containing raffinose at 26°C, then upon release from G1 the temperature was shifted up to 28°C and galactose was added to induce expression of Esp1-GFP-NLS. (Note that the cell cycle progresses slightly slower in medium with raffinose/galactose as the carbon source.) As previously described (Jensen et al. 2001), accumulation of Esp1-GFP-NLS in the nucleus occurred efficiently in the majority of cells; photomicrographs show examples of G2/M and anaphase cells with nuclear Esp1 (right) and cells not expressing Esp1-GFP (left) for comparison.