FIGURE 2.

DNA binding and self-association capacity are intact in GATA1 lacking C-terminal 95 amino acids. A, EMSA of mutant GATA1 binding to a single GATA box. Closed and open arrowheads indicate DNA and MBP fusion GATA1 complexes of wild type and mutant, respectively. Arrows indicate DNA complexes with endogenous GATA1 extracted from mouse erythroleukemia (MEL) cells. B, GST-pulldown assays performed using GST-fused GATA1 fragments consisting of the indicated amino acids. These peptides were incubated with gradated amounts of MBP fusion GATA1. The amount of captured proteins was estimated by immunoblotting to detect MBP fusion protein utilizing anti-MBP antibody (closed arrowhead). Equal volumes of the GST-fused GATA1 fragments-Sepharose bead fractions were used for the pulldown assay, and protein amounts were assessed by immunoblotting against GST (open arrowhead).