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RNF170 possesses ubiquitin ligase activity. RNF170FLAG (lanes 1–4) and *RNF170FLAG (lane 5), immunopurified from transfected HeLa cells, were incubated with E1 (UBE1), E2 (UbcH5b) and HA-ubiquitin as indicated for 30 min at 30 °C. Samples were then subjected to SDS-PAGE and were probed in immunoblots (IB) with anti-HA epitope to assess ubiquitination (upper panel) or anti-FLAG epitope to assess RNF170FLAG/*RNF170FLAG levels (lower panel).
