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Fig. 2. (A) Western blotting of membranes prepared from HEK 293 cells transiently transfected with a control plasmid or with expression vectors containing the indicated myc-tagged Ubc6 homologues. For unknown reasons the anti-myc antibody detected three bands for the epitope-tagged human homologue of the second class. (B) Immunofluorescence microscopy of HEK 293 cells expressing epitope-tagged versions of both classes of Ubc6 homologues. In the non-transfected control, anti-TRAP{alpha} was used to stain a typical ER-membrane protein. The anti-myc antibody displays a very similar staining pattern for both Ubc6 classes.