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. Author manuscript; available in PMC: 2012 Jun 14.
Published in final edited form as: Biochem J. 2010 Jul 1;429(1):127–136. doi: 10.1042/BJ20091461

Figure 6. Lysine 134 is the major site of RNF12 mediated poly-ubiquitination of LDB1.

Figure 6

HEK293 cells were transfected with expression vectors for LDB1-FLAG (WT-Ldb1 or K134R-Ldb1) and AU1-RNF12 (RNF12) as indicated. At 20 h after transfection, whole cell extracts were prepared. FLAG-tagged-LDB1 was isolated from the cell extract by immunoprecipitation with FLAG monoclonal antibody and the immunoprecipitate was analyzed by denaturing gel electrophoresis and immunoblotting with a monoclonal antibody to ubiquitin (left panel). The immunoblot was stripped and probed with antibody to LDB1 (right panel). The migration of a protein with the appropriate size for mono-ubiquitinated LDB1 is indicated with an open arrow.