Fig. 5. Foxa2 associates with Engrailed through its winged-helix/Forkhead box domain. (A) ³⁵S-labeled Foxa2 was incubated with bacterially expressed GST or GST-En2 immobilized on glutathione beads, then analysed by SDS-PAGE. Foxa2 binds to GST-En2 but not to GST. (B) Foxa2 constructs used to identify its domain(s) interacting with Engrailed. CRI to CRIV domains of Foxa2 are indicated. Numbers refer to amino acids positions. (C) Each construct was produced and labeled in vitro, and used in a pull-down experiment with GST-En2 immobilized on glutathione beads. Bound proteins were visualized after SDS-PAGE and autoradiography. All constructs containing the CRI domain of Foxa2 bind to Engrailed, and this domain (148-257) is sufficient to mediate this binding. Positions of molecular mass markers (in kDa) are indicated on the left.

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