Figure 5.

FOXC1 missense mutations that affect the DNA-binding ability of FOXC1. EMSA using the [³²P]-labeled FOXC1 site was incubated with FOXC1 wild-type and missense mutations containing COS-7 protein extracts. Recombinant FOXC1 missense proteins were equalized to recombinant wild-type FOXC1 by western blotting. The position of the FOXC1-DNA complex is indicated by the blackened arrowheads. A, FOXC1 containing the S82T missense mutation, showing reduced affinity for the FOXC1 site. FOXC1 containing the S131L missense mutations shows an ∼20-fold reduction in affinity for the FOXC1 site. B, FOXC1 containing the F112S and I126M mutations, showing DNA binding that is at wild-type or near–wild-type FOXC1-protein levels.