FIG. 2.

Fli-1 domains required for the Fli-1-GATA-1 interaction. (A) Schematic representations of the Fli-1 domains used in GST-pulldown assays. (B) Amino acid sequence of the Fli-1 Ets domain showing α-helical and β-sheet regions. (C) GST pulldowns demonstrating the Fli-1 domains required for the interaction with GATA-1. The indicated GST and GST-Fli-1 fusion constructs were expressed in E. coli strain BL-21, purified using GST-agarose beads, and incubated in the presence of ³⁵S-labeled mGATA-1. After extensive washing, the GST- or GST-Fli-1-coated beads were boiled in loading buffer and subjected to electrophoresis, after which retained GATA-1 was visualized by phosphorimaging. Lane 1 contains 10% of the input in vitro-translated ³⁵S-labeled GATA-1 protein. Other lanes contain the GST-Fli-1 deletions as shown.