Skip to main content
. Author manuscript; available in PMC: 2017 Apr 1.
Published in final edited form as: Mol Cancer Ther. 2016 Feb 16;15(4):574–582. doi: 10.1158/1535-7163.MCT-15-0467

Figure 3. CTX1 specifically targets and directly binds to HdmX.

Figure 3

A. CTX1 preferentially kills cells transformed by HdmX and not Hdm2 or shp53. IMR90 cells overexpressing the indicated constructs were treated with increasing doses of CTX1 (1, 2.5 and 5μM) and assessed for cell death at 72hr by trypan blue staining. B. CTX1 binds HdmX and to a lesser extent p53. Recombinant p53, HdmX, and/or biotin-CTX1 were incubated in vitro and streptavidin beads were used to pull down the protein complex. The bound protein was eluted and analyzed by western blot with the indicated antibodies. C. Spectral studies suggest CTX1 and HdmX directly interact. CTX1 and HdmX alone represent the spectral pattern of both agents without purification. The CTX1+HdmX sample and CTX1 Postcolumn Residual samples underwent size exclusion chromatography to remove unbound CTX1. D–E. CTX1 disrupts the interaction of recombinant HdmX/p53 but not Hdm2/p53 by ELISA and co-immunoprecipitation. The indicated drugs were used as controls. F. CTX1 disrupts the interaction of HdmX/p53 in cells. Immunoprecipitations were performed as indicated using lysate from OCI cells treated with the indicated drugs or a DMSO control. Error bars represent standard deviation.