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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 1. RNF12 has cell-specific effects on expression of LDB1 at a post-transcriptional level.

Figure 1

(A) HEK293 or MCF7 cells were transfected with an expression vector for FLAG-tagged LDB1 and either an empty expression vector (−), or vectors coding for wild type RNF12 (WT) or RNF12 with a mutation that inactivates ubiquitin ligase activity (Mut). Whole cell extracts were prepared 20 h after transfection and the extracts resolved by denaturing polyacrylamide gel electrophoresis, transferred to a membrane and then incubated with FLAG antiserum to detect epitope-tagged LDB1. A horseradish peroxidase labeled anti-rabbit secondary antibody was used with a chemiluminescent detection reagent to visualize the immunoreactive proteins. The immunoblots were stripped and then probed with antibody to ERK1 as a loading control. (B) HEK293 cells were transfected with expression vectors for LDB1-FLAG, RNF12 and empty expression vectors as indicated. At 20 h after transfection, RNA was prepared from cells. Ldb1 and GAPDH RNA were quantitated by real-time PCR. Ldb1 RNA expression in each sample was normalized to GAPDH in each sample and then relative RNA levels were normalized so that the wild type average value for the Ldb1 group was set to 1.0. Values are means +/− SEM for three separate transfections.