Figure 1

A: The T23A mutation destabilizes Cdc5. Wild-type and cdh1Δ cells were arrested in S-phase (0.2 M hydroxyurea), metaphase (5 μg/ml nocodazole) or early G1 (1 μg/ml rapamycin) for 2.5h and probed with anti-Cdc5 (Santa Cruz y300). B: Cdc5Δ cells [3] expressing Cdc5 from the indicated plasmids [16] were arrested in S-phase (0.2 M hydroxyurea), metaphase (5 μg/ml nocodazole) or early G1 (1 μg/ml rapamycin) for 2.5h. C: A full blot was probed with anti-Cdc5 to check for the possibility of proteolytic processing of cdc5^T23A. Cells were released from S-phase and rearrested at metaphase. D: Cdc5 Δ cells expressing GFP-Cdc5^WT [3] or GFP-cdc5^T23A were synchronized in S-phase, released and rearrested at metaphase. E: Cdc5-13myc cells [3] expressing Cdc5^WT or cdc5^T23A were synchronized in S-phase, released and rearrested at metaphase.