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Figure 11 | BMC Molecular Biology

Figure 11

From: Ets-2 and C/EBP-beta are important mediators of ovine trophoblast Kunitz domain protein-1 gene expression in trophoblast

Figure 11

C/EBP-β and Ets-2 interaction demonstrated by A) co-immunoprecipitation, B) and C) pull-down assays with biotinylated C/EBP oligonucleotide (-117 to -109) in JEG-3 cell extracts (CE). A) Lane 1: JEG-3 CE (positive control for western blot); lane 2: immunoprecipitation (IP) with C/EBP-β IgG (positive control for IP); lane 3: IP with Ets-2 IgG; lane 4: IP with non-immune rabbit IgG (negative control for IP). Western blot (WB) was performed with C/EBP-β IgG. B) WB performed with C/EBP-β IgG as detection reagent following pull-down assays. Lane 1: JEG-3 CE (positive control for WB); lane 2: biotinylated C/EBP oligonucleotide; lane 3: JEG-3 CE+biotinylated C/EBP oligonucleotide; lane 4: JEG-3 CE+biotinylated C/EBP oligonucleotide+250-fold molar excess unlabeled mutated C/EBP oligonucleotide; lane 5: JEG-3 CE+biotinylated C/EBP oligonucleotide+250-fold molar excess unlabeled wild type C/EBP oligonucleotide. C) WB with Ets-2 IgG as the detection agent following pull-down assays. Lane 1: JEG-3 CE (positive control for WB); lane 2: biotinylated C/EBP oligonucleotide; lane 3: JEG-3 CE+biotinylated C/EBP oligonucleotide; lane 4: JEG-3 CE+biotinylated C/EBP oligonucleotide+250-fold molar excess unlabeled wild type C/EBP oligonucleotide.

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