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. 2006 Feb 9;34(3):928–938. doi: 10.1093/nar/gkj477

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© The Author 2006. Published by Oxford University Press. All rights reserved

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ΔNp63α represses Cyclin B2, cdc2 and Topoisomerase α promoters. (A) Functional analysis of the Cyclin B2 promoters [wt and mutated in the three CCAAT boxes, see Ref. (25)] in SaoS2 cells, cotransfected with the ΔNp63α (left panel) and TAp63α (right panel) isoforms. The different AEC SAM domain mutants were cotransfected in parallel, both in the ΔN and TA configurations. (B) In the middle panels, the indicated wt and AEC ΔN and TA p63 constructs (wt, L518F and T537P) were transfected with TopoII α, wt and CCAAT-mutated promoters (left panel), or a Cdc2 promoter Luciferase plasmid (middle panel). (C) The activity of the Cyclin B2 promoter was tested with the p63 EEC C306R mutant, both in the ΔN and TA configurations. (D) The CCAAT-dependent Calreticulin promoter was cotransfected with ΔN and TA p63 expressing vectors. (E) Western blot analysis detailing the relative levels of the wt and p63 mutant proteins overexpressed, together with endogenous NF-Y to normalize protein levels.