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. 2005 Feb 14;144(7):982–993. doi: 10.1038/sj.bjp.0706127

Figure 3.

Figure 3

Effect of cyclosporin A on the interaction between CREB and its coactivator CBP as indicated by a mammalian two-hybrid assay. The luciferase reporter gene under the control of five GAL4-binding sites (5xGal4E1BLuc; 1 μg per 6-cm dish) was transfected into HIT cells together with an expression vector encoding the DNA-binding domain of GAL4 fused to the CREB interaction domain of CBP (aa 442–661) (GAL4-CBP; 1 μg per 6-cm dish). In addition, the cells were transfected with one of the following three expression vectors (2 μg per 6-cm dish): an expression vector coding for the transactivation domain of the viral protein VP16 (VP16), an expression vector encoding the CBP interaction domain of CREB-327 fused to the transactivation domain of VP16 (CREB-VP16), or an expression vector encoding a mutant CREB-VP16 protein with Ser119 of CREB mutated to Ala (CREB-S119A-VP16). Cells were treated with cyclosporin A 5 μM (CsA) as indicated. Luciferase activity is expressed relative to the activity measured in each experiment of the VP16 control. Values are means±s.e.m. (n=9).