Figure 3.

Polyubiquitinated CPY^*HA accumulates in the cytosol of the Δdsk2Δrad23 mutant. Membrane association of ubiquitinated CPY^*HA was investigated in respectively transformed wild type (WT) and Δdsk2Δrad23 strains. Microsomal extracts were separated into membrane-associated (P) and cytosolic (S) fractions. CPY^*HA was precipitated with anti-CPY antibodies and analysed by immunoblotting with anti-ubiquitin antibodies. The pellet and supernatant fractions from non-transformed strain Δdsk2Δrad23 are included to reveal the background activity of the ubiquitin antibody. Immunoblotting with HA-antibody shows localization of CPY^*HA in each fraction.