Fig. 5.
Fig. 5. HDAC3 represses GATA-2–directed transcriptional activity. / (A) The 293T cells were transfected with a luciferase reporter containing GATA consensus motif (GATA-1/Luc, 0.5 μg), together with expression plasmid for human GATA-2 (Flag–GATA-2, 0.5 μg) and human HDAC1 and HDAC3, (pcDNA3.1-HDAC1 or pcDNA3.1-HDAC3, 0.2 μg or 0.5 μg, as indicated). Luciferase activity is standardized against β-galactosidase activity from cotransfected control vector (pCMV/β-gal). The relative luciferase activities presented reflect triplicate value from a representation of at least 3 independent experiments. (B) Experiments similar to those represented in panel A were conducted using a luciferase reporter containing 2 copies of back-to-back double GATA sites from the mouse CD34 promoter (CD34 × 2/Luc., 0.5 μg; solid bar) or its mutant in which GATA sites were disrupted (mutant CD34 × 2/Luc., 0.5 μg; open bar), together with GATA-2 (Flag–GATA-2, 0.5 μg) and human HDAC3, (pcDNA3.1-HDAC3, 0.2 μg or 0.5 μg, as indicated).

HDAC3 represses GATA-2–directed transcriptional activity.

(A) The 293T cells were transfected with a luciferase reporter containing GATA consensus motif (GATA-1/Luc, 0.5 μg), together with expression plasmid for human GATA-2 (Flag–GATA-2, 0.5 μg) and human HDAC1 and HDAC3, (pcDNA3.1-HDAC1 or pcDNA3.1-HDAC3, 0.2 μg or 0.5 μg, as indicated). Luciferase activity is standardized against β-galactosidase activity from cotransfected control vector (pCMV/β-gal). The relative luciferase activities presented reflect triplicate value from a representation of at least 3 independent experiments. (B) Experiments similar to those represented in panel A were conducted using a luciferase reporter containing 2 copies of back-to-back double GATA sites from the mouse CD34 promoter (CD34 × 2/Luc., 0.5 μg; solid bar) or its mutant in which GATA sites were disrupted (mutant CD34 × 2/Luc., 0.5 μg; open bar), together with GATA-2 (Flag–GATA-2, 0.5 μg) and human HDAC3, (pcDNA3.1-HDAC3, 0.2 μg or 0.5 μg, as indicated).

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