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Figure 6


Fig. 6. Glucosidase II is co-immunoprecipitated with the Erv41p-Erv46p complex. (A) Microsomes were prepared from wild-type (FY834) and strains expressing 3HA-Erv46p (CBY770) or 3HA-Erv41p (CBY783) and solubilized with 1% digitonin. Soluble extracts were incubated in the absence (–) or presence (+) of a monoclonal antibody against the HA epitope. Precipitates were resolved on a silver stained gel. Numbers indicate bands representing Erv41p (1), 3HA-Erv41p (2) and Erv46p (3). The band indicated by and asterisk (*) was excised from a Coomassie Blue-stained gel and identified by mass spectrometry as glucosidase II. (B) Microsomes were isolated from wild-type (FY834) and Rot2p-3HA-expressing strains (SOY01117) and solubilized as above. Antibodies against the HA epitope, Erv46p or Erv41 were added as indicated at the top. Proteins eluted from the precipitate (IP), totals (T) and solubilized extracts (S) were resolved on a 12.5% polyacrylamide gel and immunoblotted with 3HA-, Erv46p-, Erv41p- and Erv25p-specific antibodies. T and S each represent 2.6% of input.





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