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. 2015 Apr 24;24(15):4185–4197. doi: 10.1093/hmg/ddv151

Figure 3.

Figure 3.

p300 interacts with ΔNp63α in human cells and catalizes in vitro acetylation of lysine K193. (A) U2OS whole cell extracts transiently co-transfected with ΔNp63α and p300 were analyzed by immunoprecipitation with an anti-p300 antibody followed by WB analysis with an anti-p63 antibody. U2OS cells, not transfected with p63 encoding plasmid were used as negative control. (B) In vitro acetylation assay performed according to the HAT assay kit protocol (Active Motif) with an H4 peptide and p53 peptides as positive controls, H4 plus anacardic acid 15 μM (an inhibitor of acetyl-transferase activity used as a negative control) and p63 peptides (peptide sequences are indicated). (C) WB analysis of U2OS whole cell extracts transiently co-transfected with ΔNp63α, ΔNp63α-K193E, ΔNp63α-K193R expression vectors (30 ng) and increasing amounts of p300 encoding plasmid (10 and 20 ng). (D) WB analysis of U2OS whole cell extracts transiently co-transfected with ΔNp63α, ΔNp63α-K193R and p300 expression vectors (30 ng and 5 ng respectively). 24 h after transfection protein half-life was measured by treating cells with 10 μg/ml of CHX.