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. Author manuscript; available in PMC: 2017 Nov 17.
Published in final edited form as: Oncogene. 2016 Sep 12;36(10):1374–1383. doi: 10.1038/onc.2016.301

Figure 5. MDM2C305F mutation attenuates p53 activation in APC loss-induced colon cancers.

Figure 5

A. Western blotting analysis of protein lysates isolated from colon (Normal) or adjacent colonic adenomas (Tumor) with three mice per genotype. p53 and MDM2 were undetectable in normal colon. The bar graph illustrates the relative protein levels in tumor samples from Apcmin/+;Mdm2+/+ and Apcmin/+;Mdm2m/m mice. Error bars, ±SEM; *P<0.05.

B. qPCR analysis of relative mRNA expression of mdm2 in colon (Normal) or colonic adenomas (Tumor) from Apcmin/+;Mdm2+/+ and Apcmin/+;Mdm2m/m mice. n=3 for each genotype. Error bars, ±SEM; NS=not significant, *P<0.05.

C. qPCR analysis of relative mRNA expression of bax in colon (Normal) or colonic adenomas (Tumor) from Apcmin/+;Mdm2+/+ and Apcmin/+;Mdm2m/m mice. n=3 for each genotype. Error bars, ±SEM; NS=not significant, **P<0.01.

D. qPCR analysis of relative mRNA expression of Apc in HCT116 cells infected with lentivirus containing either a negative control scrambled shRNA (shNC) or Apc shRNA (shApc). Error bars, ±SEM; ***P<0.001.

E. HCT116 cells were infected with lentivirus containing shRNA targeting either Apc (shApc) or a scrambled control (shNC). Following selection with puromycin, immunoprecipitation (IP) of MDM2 was performed, followed by western blotting with the indicated antibodies. Quantification of protein expression was performed using ImageJ, and the relative protein expression is indicated under the blots. The amount of RPL11 or RPL5 immunoprecipitated (relative to MDM2 IP protein levels) is also indicated under each IP blot.