Figure 4.

In vivo NF-Y–p63 interactions. (A) Endogenous NF-Y and p63 were immunoprecipitated from human primary keratinocytes with anti-NF-YC and control (anti-Flag) antbodies. Load, flow through (FT) and bound (B) materials were loaded on SDS gels and assayed in western blot analysis with the indicated antibodies. (B) Structure of the NF-YC αC structure (37). The amino acids mutated in the analysis are indicated. (C) The indicated p63-TA or ΔN- were overproduced in transfections, and the NF-YC mutants indicated were produced in Escherichia coli, together with recombinant NF-YB, and assembled. Interactions were assayed in immunoprecipitations with in vivo produced wt TAp63α (upper panels with different NF-YC mutants) or ΔNp63α (lower two panels, with NF-YB/NF-YC and the NF-YC I115P mutant).