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. 2002 Oct;22(20):7120–7133. doi: 10.1128/MCB.22.20.7120-7133.2002

FIG. 6.

FIG. 6.

Mapping of the Pitx1 domains, which interact with IRF3 or IRF7. (A) Interactions of IRF3 or IRF7 with Pitx1 were examined in pull-down assays, using MBP fusion proteins: MBP-Pitx1, MBP-HD (residues 84 to 153), MBP−N+HD (residues 1 to 153) and MBP-LacZ as a control. Proteins were incubated in the presence of in vitro-translated 35S-labeled IRF3 and IRF7 proteins as described for Fig. 3. (B) Pitx1 lacking the HD (Pitx1-ΔC6) protein was labeled by in vitro translation and tested for binding to MBP-IRF3, MBP-IRF7, or MBP-LacZ. (C) The interaction of endogenous IRF3 or IRF7 protein with Pitx1 was examined by incubating MBP fusion proteins (MBP-Pitx1, MBP-HD, MBP-N+HD, and MBP-LacZ) in the presence of nuclear extracts from NDV-induced cells. Binding of IRF3 or IRF7 to Pitx1 was analyzed by Western blotting using an anti-IRF3 or anti-IRF7 antibody. The HD is the region of Pitx1 that interacts with IRF3 and IRF7.