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. 2003 Mar 17;160(6):857–874. doi: 10.1083/jcb.200209097

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Copyright © 2003, The Rockefeller University Press

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Figure 2. — GAL-CLA4t expression suppresses the spindle checkpoint defects of mad2 and bub2 mutants. (A) Cultures of wild-type, bub2Δ, mad2Δ, and mad2Δ bub2Δ mutants, either lacking (W303, ySP3138, ySP1070, and ySP1086) or carrying one integrated copy of GAL-CLA4t (ySP2622, ySP2626, ySP2752, and ySP3068) were arrested in G1 by α-factor treatment and then released into YEPRG medium containing nocodazole (t = 0). (B) The same strains and procedure as in A were used, but 10 μg/ml α-factor was re-added to all cultures at t = 100' after release (at least 90% of budded cells) to prevent them from entering a new cell cycle. At the indicated times cells were collected for FACS^® analysis of DNA contents (A) and for Western blot analysis of Clb2 and Sic1 protein levels (B). Swi6 was used as loading control. Cyc, cycling cells.

Figure 2. — GAL-CLA4t expression suppresses the spindle checkpoint defects of mad2 and bub2 mutants. (A) Cultures of wild-type, bub2Δ, mad2Δ, and mad2Δ bub2Δ mutants, either lacking (W303, ySP3138, ySP1070, and ySP1086) or carrying one integrated copy of GAL-CLA4t (ySP2622, ySP2626, ySP2752, and ySP3068) were arrested in G1 by α-factor treatment and then released into YEPRG medium containing nocodazole (t = 0). (B) The same strains and procedure as in A were used, but 10 μg/ml α-factor was re-added to all cultures at t = 100' after release (at least 90% of budded cells) to prevent them from entering a new cell cycle. At the indicated times cells were collected for FACS^® analysis of DNA contents (A) and for Western blot analysis of Clb2 and Sic1 protein levels (B). Swi6 was used as loading control. Cyc, cycling cells.