SHP does not interfere p53 and Mdm2 interaction.

<p>A–B: GST pull down to determine the <i>in vitro</i> interaction of SHP with p53. Flag-p53 was <i>in vitro</i> translated and used to interact with various GST-SHP fusion proteins, including GST-FL (full length), GST-2 (interaction domain), GST-1+2 (repression domain deletion), GST-3 (repression domain), and GST-2+3 (N-terminal domain deletion), that were expressed from bacterial Escherichia coli BL21/DE3/RIL. C: Immunoprecipitation and Western blots. Plasmids expressing Myc-p53 were cotransfected with Flag-SHPWT, Flag-SHP38H, Flag-SHP170N, and Flag-SHP171A plasmids in Hela cells. Co-IP and WB were performed with corresponding antibodies as indicated. D–E: Western blots to determine the effect of SHP on p53 and Mdm2 interaction. D: HA-Mdm2 (2 µg), Myc-p53 (4 µg) and Flag-SHP (2 µg) plasmids were cotransfected in 293T cells, and their proteins were detected using anti-HA, anti-Myc and anti-Flag antibodies, respectively. E: HA-Mdm2 (2 µg), Flag-p53 (4 µg) and Flag-SHP (2 µg) plasmids were cotransfected in Hela cells, and their proteins were detected using anti-HA and anti-Flag antibodies, respectively.