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Levels of ubiquitin in doa4Δ cells carrying the ubiquitin-encoding YEp96 multicopy plasmid. Anti-ubiquitin immunoblot analysis of logarithmic phase extracts from wild-type (MHY501) and doa4Δ (MHY623) cells cotransformed with plasmid-borne Deg1-Ura3 and either pES12 (vector) or YEp96 (CUP1UB). The same transformants were used for the pulse-chase analysis in Figure 4, B–D. [125I]protein A was used to detect ubiquitin and Pgk1 levels.
