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Vacuole targeted H+-ATPase mutant has a low affinity for rafts. WT (L3852) and pma1-7 (ACY7) cells were radiolabeled for 10 min at 24°C and chased for 30 min at the same temperature. After TX100 extraction (1% at 4°C) and density gradient centrifugation the ATPase was immunoprecipitated from detergent resistant (R) and soluble (S) fractions and analyzed by SDS-PAGE and phosphorimaging.
