Figure 5.

Sucrose density gradient fractionation of cells (YTS3) expressing Ha-tagged Gid2p from the chromosome. After 16 h of growth on ethanol medium, the cells were shifted to glucose medium. The cells were analyzed before (A) and 30 min after shift to glucose medium (B). From the sucrose gradient, fractions were collected from the top of the gradient (fraction 1) and analyzed by immunoblotting with antibodies against Ha (Ha-tagged Gid2p), cytosolic PGK, Fas (cytosolic fatty acid synthase), CPY (vacuolar carboxypeptidase yscY); vacuolar API, and ER lumenal Kar2p. The fractions were further tested for their Guanosine diphosphatase (Golgi) activity. Further details are given in the text.