Interaction of OKL38 and p53 proteins.

<p>(A) p53 was co-immunoprecipitated by the M2 agarose beads together with the FLAG-OKL38 expressed in p53^+/+ HCT116 cells, but not from the control cells without FLAG-OKL38 expression. (B) Nuclear extracts of U2OS cells were precipitated with normal mouse IgG or anti-p53 antibody and blotted with anti-p53 and anti-OKL38 antibodies. (C) Nuclear extracts of U2OS cells were precipitated with normal rabbit IgG or anti-OKL38 antibody and blotted with the anti-OKL38 and anti-p53 antibodies. Arrow denotes the position of p53. (D) GST-pull down experiments showed that GST-OKL38 beads retained p53 from nuclear extracts. (E) Similarly, GST-p53 beads retained FLAG-OKL38 expressed in Hela cells. (F) Illustration of OKL38 domain structures (TrkA and Pyr_Redox_2 domains) and the GST-truncation OKL38 constructs used to analyze OKL38 and p53 interaction. (G) Interaction of p53 with GST-OKL38 full length (residues 1–477) or its truncation derivatives in GST-pull down assays. Different percentages of input were loaded to serve as a control. (H) Illustration of p53 domain structures and its derivatives used in studying p53 and OKL38 interaction. (I) GST-pull down assays to detect interaction of FLAG-OKL38 with GST-p53 and its truncation derivatives.