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. 2018 Mar 25;27(5):976–986. doi: 10.1002/pro.3405

Figure 2.

Figure 2

p53 enhances the interaction between HDMX and HDM2. (A) ELISA assay showing the effect of p53 on the HDMX‐HDM2 interaction. The plates were covered with a fixed amount of HDMX (10 ng/μL) and increasing amounts of HDM2 (0–10 ng/μL) in the presence or absence of a fixed amount of recombinant purified p53 (10 ng/μL). The interaction between HDMX and HDM2 increases in the presence of p53. Anti‐HDM2 YB25 antibody was used to develop the assay. The average of five independent experiments is shown. (B) ELISA assay showing that HDM2 interacts with p53 in a specific manner; HSP90 was used as a negative control of the interaction. The plates were covered with a fixed amount of p53 or HSP90 (20 ng/μL) and increasing amounts of HDM2 (0–20 ng/μL) and developed with 2A10 anti‐HDM2 antibody. (C) Coimmunoprecipitation assay performed in the H1299 cell line. Cells were transfected with HDMX and HDM2, with or without p53. The 4B2 anti‐HDM2 antibody was used to pull down HDM2, and the ChX chicken anti‐HDMX antibody was used to develop the western blot.