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Pulse-chase fractionation of membranes from a end4–1 chs6 double mutant. Cells were incubated at 37°C for 10 min, pulse-labeled with 35S-Promix for 10 min, and chased for 30 min. Membranes were separated by differential centrifugation (Con A fractionation) and proteins were evaluated by immunoprecipitation, SDS-PAGE, and PhosphorImager analysis. P indicates the 10,000 × g pellet, and S indicates the 10,000 × g supernatant.
