FIGURE 2:

Failure to export Chs2 from the ER in the cdc14-NES mutant can be bypassed by abolishing the Cdk1 sites on Chs2. cdc14-NES SEC63-CFP containing GAL-CHS2-YFP (A) or GAL-CHS2(4S-to-A)-YFP (B) cells was arrested in YP/Raff with Noc for 5 h, following which CHS2 was induced with 2% Gal for 45 min. The cells were then shifted to 37°C for 15 min before release into YPD prewarmed at 37°C (ii). At 30 min after release into YPD, 32 μM Lat B was added into both cultures and cells further incubated at 37°C for an additional 90 min. (A, iii; B, iii) Chs2-YFP signals were enumerated as in Figure 1.