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Mutations in specific ubiquitin-conjugating enzymes do not inhibit ERAD in vitro. Microsomes and cytosol from either the ubc6 ubc7 (ubc) or an isogenic wild-type strain (WT) were used in an in vitro ERAD assay as previously described (8). Addition of ubiquitin-aldehyde (UA) to wild-type cytosol to a final concentration of 1 μM, did not inhibit degradation of pαF in vitro. Results represent the means of at least two independent determinations.
