FIG. 4.

RARα can be recruited to GATA motifs in DNA through interaction with GATA-2. 293T cells were transfected with the expression plasmids encoding RARα (A) or RXRα (B) and Flag-tagged GATA-2 as indicated. Nuclear extracts of the cells were then prepared and incubated with biotinylated oligonucleotides harboring GATA motifs (wild-type [wt] GATA oligonucleotides) or biotinylated mutant oligonucleotides in which GATA motifs were changed to TTTA (mutant [mt] GATA oligonucleotides). The oligonucleotides were then recovered by streptavidin-agarose beads, and the copurified proteins were analyzed by Western blotting with anti-RARα or anti-RXRα (top) and anti-Flag (bottom) antibodies. Input (10% input) was used as a control. (C) Nuclear extracts of human myeloid KG-1 cells were incubated with biotinylated oligonucleotides harboring GATA motifs and pulled down with streptavidin-agarose beads. The pulled-down materials were analyzed by Western blotting with antibodies against GATA-2, RARα, and RXRα.