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. 2007 Jul 23;27(19):6818–6831. doi: 10.1128/MCB.00375-07

FIG. 3.

FIG. 3.

TRB3 inhibits C/EBPβ transcriptional activity. (A) Characterization of the ability of the exogenous C/EBPβ to induce PPARγ in fibroblastic cells Swiss 3T3 in the presence or absence of TRB3. A total of 40 μg of total protein extracts was loaded, and adipocyte differentiation marker expression was assessed by Western blotting. The results are representative of two independent experiments. (B) Coimmunoprecipitation experiments of C/EBPβ with TRB3 or a N-terminal truncated form of TRB3. A total of 500 μg of protein was used to immunoprecipitate FLAG-tagged TRB3 or FLAG-tagged ΔTRB3. The elution was loaded equally on two gels to assess TRB3 and C/EBPβ protein amounts. The results are representative of two independent experiments. (C) Schematic of the structures of the C/EBPβ mutant isoforms used in the deletion analysis. TAD, transactivation domain; RD1, repression domain 1; RD2, repression domain 2; DBD, DNA-binding domain; LZ, leucine zipper. (D) Coimmunoprecipitation experiments of deletion mutants of C/EBPβ isoforms with TRB3. A total of 250 μg of protein were used to perform the different immunoprecipitations. The elutions were analyzed by Western blotting to assess TRB3 and C/EBPβ isoform protein amounts.