Effect of USP7 on the DNA binding activity of p53 in vitro.

<p>(A) Schematic representation of the p53 proteins used in this study showing the transactivation (Trans), DNA binding core, tetramerization (Tet) and USP7-binding and regulatory (USP7/Reg) regions. (B) EMSA showing titration of latent p53_82–393 in the presence of 20 µM of BSA negative control (lanes 2–5) or 20 µM USP7 (lanes 6–9). (C) EMSA performed with fixed amount (12 µM) of p53_82–393 and with 5 µM, 10 µM or 20 µM of USP7 (lanes 3–5) or BSA (lanes 6–8). Incubation of 20 µM of USP7 alone or BSA alone with labeled probe in the absence of p53 is also shown (lanes 9 and 10). (D) EMSA showing titration of active p53_82–360, in the absence (lanes 2–5) or presence of USP7 (lanes 6–9). Quantification of the discreet shifted bands for parts B,C and D are shown in the graphs, with USP7 in black and BSA in grey.