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Co-transfection of ISG15 with mutant forms of the ISG15 conjugating enzymes. HEK293T cells were transfected with plasmids expressing the indicated wild-type or mutant proteins. The plasmid expressing ISG15 contains an N-terminal 3×FLAG tag and the plasmid expressing Herc5 contains an N-terminal TAP tag. Cell extracts were prepared 24 h posttransfection and analyzed by immunoblotting with an antibody against the FLAG tag.
