Figure 5.

Late mitotic release of Cdc14 coincides with Brn1 relocalization. (A) Micrographs showing IF for tagged Brn1 and Cdc14 performed on exponentially growing cultures of strains GA-3600 (CDC14-3HA, BRN1-13MYC, TUB1-GFP) and GA-3601 (lte1Δ, CDC14-3HA, BRN1-13MYC, TUB1-GFP). Tubulin-GFP and DAPI are visualized by direct epifluorescence. Cultures were grown at 30°C and fixed for IF or incubated for 2 h at 25°C before fixation. Phenotypes shown in panels a, b represent 80% of the wt phenotypes, whereas panels c, d represent 60% of the lte1 cells when grown at 25°C. A scheme shows dynamics of Cdc14, spindle and Brn1 from G2/M to telophase in wt or lte1 mutants. (B) Strain GA-3819 (cdc14-1 BRN1-GFP, CFP-TUB1) was grown at 23°C. On top, a scheme of the experiment, and below, the frequency of compact Brn1 spirals in wt and mutant strains after nocodazole block and cdc14-1 inactivation. A representative micrograph shows cdc14-1 mutant cells after 90 min release at 37°C, with Brn1-GFP (green) and CFP-Tub1 (red). Bar=5 μm.