FIG. 3.

Functional and biochemical characterization of C/EBPβ-interacting domains of TIF1β. (A) (Upper panel) Schematic representation of several TIF1β expression vectors; numbers denote amino acid positions. (Lower panel) Transient transfection assays. BHK cells (in 3.5-cm-diameter petri dishes) were transfected with 0.5 μg of AGP (WT)-CAT, 0.1 μg of pCMV-C/EBPβ, and 2 μg of pcDNA3-TIF1β (full length [fl] or from amino acid 1 to 563 or 1 to 372, or full length but with amino acids 80 to 383 deleted). (B) (Upper panel) Schematic representation of several GST-TIF1β fusion proteins. (Lower panel) Protein pull-down assay. Glutathione bead-immobilized recombinant GST-TIF1β (fl or from amino acid 80 to 383, 383 to 563, or 383 to 834) incubated with full-length recombinant C/EBPβ (100 ng). After extensive washes, the protein complex was analyzed by immunoblotting with anti-C/EBPβ antibody. −, pcDNA3 vector control.