PLZF-RARα has a capacity to modulate GATA-2 activity.

Luciferase reporter gene assays using 293T cells were conducted as described in the legend to Figure 2. (A) Expression plasmids for GATA-2 (GATA-2/pMT2, 100 ng), PLZF (PLZF/pSG5; 500 ng), PLZF-RARα (PLZF-RARα/pSG5, 500 ng), and PML-RARα (PML-RARα/pMT2, 500 ng) were used as indicated, in combination with GATA-1/Luc or mutant GATA-1/Luc reporter plasmid (0.5 μg). ATRA (RA; 1 μΜ) (solid bar) or solvent (dimethyl sulfoxide) (open bar) was added to the culture media 24 hours after transfection, and luciferase activities were measured 24 hours later. (B) Expression plasmids for GATA-2 (GATA-2/pMT2, 100 ng) and PLZF-RARα (PLZF-RARα/pSG5, 500 ng) were used in combination with CD34 × 2/Luc or mutant CD34 × 2/Luc reporter plasmids (0.5 μg), as indicated. ATRA (RA; 1 μM) trichostatin A (TSA, 100 nM), or both, was added to the culture media 24 hours after transfection, and luciferase activities were measured 24 hours later. Luciferase activities were normalized as described in the legend to Figure 2 and were presented as fold increase in activity from the wild-type GATA-1/Luc (A) or CD34 × 2/Luc reporter (B) alone in the absence of the chemical reagents.