Figure 5.
GFP-Ccc1 and GFP-NtΔCcc1 protein stability in normal and high-iron conditions. Yeast ccc1Δ cells transformed with either pUG36-CCC1 (PMET17-GFP-CCC1) or pUG36-NtΔCCC1 (PMET17-GFP-NtΔCCC1) plasmids were cultivated in SC-Ura-Met to exponential phase and 3 mM FAS was added (or not) for 1 h. Expression was stopped by addition of 3 mM methionine, and aliquots were isolated at the indicated time points. Total proteins were extracted, and Ccc1 protein levels were determined by immunoblotting with anti-GFP antibody. Equal amounts of total proteins were loaded in each lane. Three independent biological experiments were performed, and GFP-Ccc1 and GFP-NtΔCcc1 protein half-lives was determined. The average half-life and its standard deviation are presented.