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. Author manuscript; available in PMC: 2014 Jan 21.
Published in final edited form as: Curr Biol. 2012 Dec 27;23(2):162–167. doi: 10.1016/j.cub.2012.11.049

Figure 4. Elmo1 promotes the ubiquitination of Med31.

Figure 4

A. Elmo1 promotes the mono- and di-ubiquitination of Med31. Immunoblotting of GST precipitates from cell lysates expressing HA-Ubiquitin, GST-Med31, and Elmo1-GFP (wild type, PxxP) or Elmo1 AxxA-GFP (mutant). Molecular weight makers are indicated. B. Wild type Elmo1 promotes the cytoplasmic mono-ubiquitination of Med31. Immunoblotting of GST precipitations from cytoplasmic and nuclear extracts of cells expressing HA-Ubiquitin, GST-Med31, and Elmo1-FLAG, or Elmo1 truncation mutants 1–558 and 532–727. Mono-ubiquitinated Med31-Ub was primarily found within the cytoplasmic extract, while Med31-(Ub)2 was observed within both cytoplasmic and nuclear extracts. Coexpression of Med31 with either full length (1–727) or PxxP-containing fragment (532–727) of Elmo1 enhanced the levels of both cytoplasmic Med31-Ub and nuclear Med31-(Ub)2 compared to coexpression with a PxxP-deletion mutant of Elmo1 (1–558).