FIG. 3.

Effects of E1A and VP16 on Eya1-Dach1 synergistic activation. (A) One-half microgram of reporter, pGLMRG5, was cotransfected with or without 0.3 μg of pfDach1 and with 0.3 μg of pMEya1 plasmids. Increasing amounts of E1A and its deletion mutation E1Adelta CR1, denoted as E1Amut (5, 50, and 500 ng), were cotransfected. Relative luciferase activity (Act.) was normalized as the protein concentration of each nuclear extract, because E1A influenced the β-galactosidase activity of the internal control pEFBOSβ-GAL. The activity in the presence of pMEya1 and pfDach1 was set at 100. (B) One-half microgram of reporter pGLMRG5 was cotransfected with or without 0.3 μg of pfDach1 and with 0.3 μg of pMEya1 plasmids. Increasing amounts of pVP16-N (4, 40, and 400 ng) were cotransfected. The activity in the presence of pMEya1 and pfDach1 was set at 100. (C) The reporter pGLMRG5 (0.3 μg) was cotransfected with increasing amounts of pCMVHACBP (0.083, 0.25, and 0.75 μg) in the presence of 0.15 μg of pMEya1 and 0.15 μg of pfDach1 (columns 2 to 13) in the presence of 0.01 μg of E1A or E1Amut (columns 6 to 9 or 10 to 13). The luciferase activity in the presence of pMEya1 and pfDach1 (column 2) was set at 1. (D) pGLMRG5 and pFA-CHOP were cotransfected with increasing amounts of E1A and its deletion mutation E1Amut (5, 50, and 500 ng). The luciferase activity in the presence of pFA-CHOP was set at 100.