FIGURE 4:

(A) Cdc14 nucleolar release precedes Chs2 neck localization. CHS2-YFP CDC14-ECFP cells were arrested in YPD/Noc at 24°C. After 4 h, the cells were then released from Noc arrest and mounted onto an agarose pad for time-lapsed imaging. Images were acquired at 3-min intervals, with nine Z-planes at 0.5 μm between each plane for Cdc14-ECFP (i) and single planes for Chs2-YFP (ii). Merged images are shown in (iii). (B) Premature Cdc14 dispersal and Chs2 ER export from nucleolus at metaphase can be triggered by Sic1 overexpression. CDC14-ECFP CHS2-YFP NET1-mCHERRY GAL-SIC1-myc(4copies) cells were arrested in Noc in YP/Raff for 5 h at 24°C. At Noc arrest, Glu (i) or Gal (iii) was added to the cells to a final concentration of 2%, and cells were mounted onto agarose pads containing Noc and examined by time-lapsed microscopy. Images were acquired as in A but only selected time points are shown. In addition, Net1-mCherry signals were captured similar to Cdc14-ECFP. Percentages of cells with Cdc14-ECFP nucleolar localization or dispersal and Chs2-YFP neck localization were plotted (ii, v).