Fig. 6. Analysis of the effects of Cdc2p-as inactivation upon the regulation of cytokinesis. (A) Small cells of the strain cdc2-as mob1-GFP pREP3X-mad2 were selected by centrifugal elutriation. A sample was taken when cells were in mitosis, and analogue or DMSO was added aliquots of the remaining cells. Ten minutes later, cells were harvested, fixed and stained to examine -tubulin and GFP by indirect immunofluorescence. The images show representative DAPI staining, -tubulin and GFP indirect immunofluorescence in the three samples. For the T10 analogue panels, the left-hand image shows a cell with a single Mob1p-GFP ring, whereas the right hand images show a cell with a double ring. Note that Mob1p-GFP is present on the SPBs throughout mitosis. Quantification is presented in Table 2. (B) cdc2-as cdc13-GFP pREP3X-mad2 cells were treated as described in the legend for panel A. (C) Cells of the strain cdc2-as cdc7p-GFP pREP3X-mad2 were treated as described in the legend for panel A. A sample was examined 20 minutes after addition of analogue. Cells were fixed and stained with DAPI and Calcofluor. Scale bars: 10 µm.