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ISG15 conjugation to exogenously expressed targets is enhanced by the co-expression of Herc5. HEK293T cells were transfected with a plasmid expressing an epitope-tagged target protein either alone (lane 1), or combined with Ube1L, UbcH8 and ISG15 (lane 2), or combined with Ube1L, UbcH8, ISG15 and Herc5 (lane 3). Cell extracts were prepared 48 hours posttransfection and analyzed by immunoblotting using the indicated antibodies.
