Figure 9

Extracellular proteins involved in different functions are present in the vesicle-enriched fraction. Wild-type cells expressing GFP tagged proteins were grown in YPKG for 3d and extracted. Total extracts were centrifuged first at 3,000 × g for 10 min and 200,000 × g for 2 hours. The 200,000 × g pellet fraction was resuspended in PBS and incubated in the absence or presence of 2% SDS for 30 min. Following incubation, samples were re-centrifuged at 200,000 × g for 2 hours. The distribution of proteins in the 200,000 × g supernatant (S) and pellet (P) fractions was determined by Western blotting.