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Fig. 1. Activation of new growth zones in bipolar G2 cells. (A) Branching cells during cdc25-22 block. Cells were blocked at 36.5°C for 1, 2 and 3 hours, prior to MBC/DMSO addition (length of block prior to MBC/DMSO addition shown in parentheses in the graph). Black arrows indicate time of drug treatment. The percentage of branched cells is an average of three experiments (unless otherwise stated) with s.e.m. values smaller than 5%. 100-200 cells were counted at each time point. (B) Brightfield images of blocked (3 hours) cells treated for 3 hours with DMSO (top left) or with 50 µg/ml MBC (top right and lower panels). Staining with TAT-1 antibody of MBC-treated cells, treated as above (bottom). (C) Cdc25-22 blocked (3 hours) cells treated with DMSO (left), MBC for 1 hour (middle) and for 3 hours (right), stained for actin (upper panel), with DAPI (middle panel) and with calcofluor (lower panel). (D) Frames from movie of cdc25-22 cdc13-YFP strains blocked at 36.5°C for 3 hours and then treated with MBC at t=0, at start of time-lapse. Frames were taken every 10 minutes, using an Axioplan Zeiss microscope. Bars, 5 µm.
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