Figure 6.

Spc72-GFP and DNA localization in cnm67Δ cells. (A) In small budded cells before or after SPB separation, the Spc72-GFP signal localizes to bright spots close to the nuclear periphery corresponding to the SPBs. (B) During later cell cycle stages most Spc72-GFP still localizes to the nuclear periphery, but spots of Spc72-GFP signal can be detected in the bud not associated with the nuclear periphery. This is obvious in a binucleate anaphase cell with two buds (right panel). The larger bud from the previous cell cycle does not contain DNA (consistent with two anaphase nuclei in the mother cell), but Spc72-GFP can be detected in this bud. Also, the new bud already shows Spc72-GFP spots not associated with DNA. The DNA was stained with DAPI. These still images consist of five FITC z-axis planes (green), one DAPI plane (blue), and one phase-contrast image (red) merged into one plane. Bar, 10 μm. Movie 7: γ-Tubulin complex detachment as seen in Spc72-GFP–labeled cnm67Δ cells followed for 15 min. Acquisition interval, 15 s; movie speed, 10 frames per second = 2.5 min/s. Three z-axis plane fluorescence images were acquired and merged.