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Figure 8


Fig. 8. PKC{alpha}-depleted ER and cytosol has no effect on PCTV budding. (A1) Cytosol was twice treated either with bead-bound IgG (Cyto) or bead-bound anti-PKC{alpha} antibodies (PKC{alpha}-dep Cyto). 30 µg of treated cytosol protein were separated by 12% SDS-PAGE, transferred to a nitrocellulose membrane and PKC{alpha} was identified by immunoblotting (ECL). (A2) ER was treated either with cold HEPES (ER) or cold HEPES plus 2 M urea (2 M Urea washed ER). PKC{alpha} was detected by immunoblot (ECL). (B) ER was washed with 10 mM HEPESand cytosol was treated with bead-bound IgG (IgG treated). PKC{alpha} was removed from cytosol by immunodepletion (Fig. 8A1) and ER by 2 M urea wash (Fig. 8A2). Treated ER (500 µg) and cytosol (1 mg, PKC{alpha} depleted) were then used in a PCTV-budding assay and the results shown. Data are the mean ± s.e.m.