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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 2. A slowly migrating form of LDB can be identified following expression of RNF12 and LDB1 in HEK293 cells.

Figure 2

HEK293 cells were transfected with vectors for LDB1, FLAG-tagged wild type (WT) or mutant (Mut) RNF12 or empty expression vector (−) as indicated. At 20 h after transfection, whole cell extracts were prepared. A portion of the whole cell extract was analyzed by denaturing gel electrophoresis and immunoblotting with polyclonal antisera to LDB1 to detect the input expression of the LDB1 (5% Input, lanes 1–4). FLAG-tagged-RNF12 was isolated from the cell extract by immunoprecipitation with FLAG monoclonal antibody and the immunoprecipitate was analyzed by denaturing gel electrophoresis and immunoblotting with LDB1 rabbit polyclonal antibody to detect co-immunoprecipitated LDB1 (Immunoprecipitate, lanes 5–8). Immunoblotting detected proteins with the appropriate migration for full-length LDB1 and a more slowly migrating form of the protein (open arrow).