Fig. 6. (A) Wild-type and clp1 cells carrying integrated copies of Sid1p-GFP and Cdc7p-GFP were cultured to exponential growth phase at 24°C, synchronized in early G2 by centrifugal elutriation and then treated with a low dose (0.2 µM) of Lat A or DMSO (solvent control) and cultured at 32°C. Cells were subsequently fixed at 30 minute intervals, stained with antibodies against GFP (see Materials and Methods), and scored for localization of Sid1p-GFP (left) and Cdc7p-GFP (right) to the SPB. (B) Wild-type and cdc14-118 cells carrying an integrated copy of Clp1p-GFP were treated with 0.2 µM Lat A or DMSO, shifted to 32°C for 3.5 hours, fixed and then stained with DAPI. Clp1p-GFP localization was monitored using GFP autofluorescence.