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. 2009 May 5;37(12):3996–4009. doi: 10.1093/nar/gkp332

Figure 1.

Figure 1.

Two-hybrid screening identifies FHL2 as an Id2-interacting protein. (A) The left panel shows the schematic illustration of DBD–Id2–DBM–δHLH bait protein. The mutant D-box was described in ref. (22). The right bargraph shows the results of β-galactosidase activity assays. Yeast AH109 cells were transformed with the indicated GAL4–DBD and GAL4–AD chimeric constructs, and the β-galactosidase activity was measured by a liquid O-nitrophenyl-β-d-galactoside assay. The experiment was repeated twice, and three different yeast transformants were used for each measurement. Control represents the interaction of p53 with SV40 large T-antigen proteins (#P < 0.01). (B) Mammalian two-hybrid assay was performed with 293T and MCF-7 cells, in which pG5-Luc was cotransfected with the indicated vectors. Approximately 42 h after transfection, the cells were harvested and luciferase activity was measured. The relative luciferase activity levels (normalized) for pACT and pBIND parent vector transfections were arbitrarily assigned a value of 1. All experiments were performed in triplicate and were repeated at least three times, and the results are expressed as mean ± SEM.