Fig. 6. Identification of the domains of Engrailed interacting with Foxa2. (A) Engrailed truncated constructs produced as GST fusions to identify Foxa2-binding domains within the En2 sequence. The homeodomain (EH4), as well as other previously characterized domains of Engrailed [EH1-5 (Duboule, 1994)] are indicated. Numbers refer to amino acids positions. (B) ³⁵S-labeled Foxa2 was incubated with immobilized GST-fusion proteins. Bound radioactive protein was visualized, following SDS-PAGE, using autoradiography. Left panel illustrates that the homeodomain and the 146-199 domain fully retain Foxa2 (compare with input). Right panel demonstrates that, compared with the full 146-199 domain, the two sub-domains 146-167 and 168-199 bind Foxa2 poorly.

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