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E3 Ubiquitin Ligase Activity of DNF.
GST-DNF was expressed and purified from E. coli and tested for ubiquitination activity in the presence of yeast E1, human E2 (Hubc5b or Hubc5a), and ubiquitin. The immunoblots were probed with anti-Ub antibodies (top panel) to detect ubiquitinated E. coli proteins. Anti-GST antibodies (bottom panel) were used to detect GST-DNF.
