Degradation of MCC transporters is accelerated in mutants affected in the domain formation. (A and B) Exponentially growing cultures of wild-type (WT), nce102Δ, and pil1Δ cells expressing Can1-GFP (A) and Fur4-GFP (B) were treated with cycloheximide. At the given time points, total membranes were isolated from the culture aliquots (see Materials and methods). The membrane proteins were resolved by SDS-PAGE, and Can1-GFP and Fur4-GFP were detected by anti-GFP antibody on Western blots. 2.5 μg of the total protein was loaded into each lane. Black lines indicate that intervening lanes have been spliced out.