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. 2002 Oct 7;99(22):14298–14302. doi: 10.1073/pnas.162491399

Fig 4.

Fig 4.

Effect of cisplatin on yeast Ctr1 protein. (A) Degradation of yeast Ctr1p upon cisplatin treatment. The Ctr1 protein was tagged at its C terminus with an HA epitope by modification of the genomic CTR1 locus (YSI37). Cycloheximide (100 μg/ml) was added to the CTR1-HA cells 30 min before cisplatin treatment to block new protein synthesis. Cells were then exposed to 0.1 mM or no cisplatin, and samples were taken at the indicated times. The level of Ctr1 protein was determined by Western blotting using anti-HA antibodies. The loading of proteins in each lane appeared equal by Coomassie blue staining. (B) Localization of yeast Ctr1p after cisplatin or copper treatment. The Ctr1 protein was tagged at its C terminus with GFP by modification of the genomic CTR1 locus (YSI38). Cells were incubated with 100 μg/ml cycloheximide for 30 min to block new protein synthesis, treated with 1 mM cisplatin for 4 h, and analyzed by fluorescence microscopy.