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Fig. 5. Encore associates with Cyclin E, Cul1, the proteasome 19S-RP subunit S1 and
the proteasome 20S-CP LMP7. (A) Immunoblot (IB) using antibodies against
Cyclin E and Ubiquitin showing that Cul1 and Encore can immunoprecipitate
Cyclin E-Ubn. Immunoprecipitation with S1 and LMP7 precipitates
several polyubiquitinated Cyclin E forms. The control lane (co) refers to
immunoprecipitations that use unrelated antibodies raised in the same animal
as the antibody used for the immunoprecipitation test. (B) IB using antibodies
against Encore shows that Cyclin E, Cul1, S1 and LMP7 associate with Encore in
the ovary. Notice that the amounts of immunoprecipitated protein in the Cul1
panel are comparable in wild-type and encore mutant extracts. The S1
and LMP7 immunoprecipitation lanes show that several forms of Encore
associates with S1 and LMP7. The wild-type extract lane indicates the position
of Encore in the gel. The extracts were run in 7%Tris-Acetate Nupage gel
(Invitrogen).
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