FIGURE 4.

AEC-associated, SAM domain, p63α mutations increase binding to SATB2. A, mutations associated with EEC syndrome are found exclusively in the DNA binding domain of p63 whereas those associated with AEC syndrome map to the SAM domain of p63α. A nonsense mutation leading to a premature stop codon upstream of the SAM domain has also been associated with AEC syndrome. B–D, various wild-type or mutant 3×FLAG-ΔNp63α constructs were co-transfected with T7-SATB2, and lysates were immunoprecipitated (IP) with anti-FLAG antibodies. Bound proteins were separated on SDS-PAGE and analyzed by immunoblotting (IB). E, wild-type 3F-ΔNp63α or 3F-ΔNp63α mutants were in vitro translated together with wild-type T7-ΔNp63α and immunoprecipitated with anti-FLAG antibodies. Bound proteins were resolved on SDS-PAGE and immunoblotted with anti-FLAG or anti-T7 antibodies. F and G, HEK293 cells were co-transfected with the indicated expression plasmids. Lysates were immunoprecipitated with control (anti-HA) or anti-FLAG antibodies and blotted with anti-FLAG or anti-T7 antibodies.