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. 2010 Mar 31;30(11):2750–2761. doi: 10.1128/MCB.00804-09

FIG. 2.

FIG. 2.

Identification of a p400 fragment that binds to TIP60 and suppresses TIP60-enhanced basal p21 promoter activity. (A) Schematic representation of the p400 polypeptide. The p400 fragments (F1 to F6) used for coimmunoprecipitation mapping and luciferase reporter assays are shown. (B) Association of p400 fragments with TIP60 in 293T cells. HA-TIP60 and the indicated Flag-p400 fragments were coexpressed in 293T cells, and after immunopurification on M2 agarose, bound Flag-p400 fragments and associated HA-TIP60 were monitored by immunoblots with anti-Flag (middle) and anti-HA (bottom) antibodies, respectively. (C) Effect of p400 fragments on TIP60-enhanced p21 promoter-driven luciferase activity. HCT116 cells were cotransfected with a p21 promoter-driven reporter plasmid (pWWP-luc) and plasmids encoding the indicated p400 fragments in the presence or absence of ectopic TIP60 expression. Plasmids expressing wild-type TIP60 (left) or HAT mutant TIP60 (right) were used for ectopic TIP60 expression. After normalization to an internal control reporter, the luciferase activity obtained from a vector control was set to 100, and relative values of each p400 fragment were then calculated.