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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Mol Cancer Res. 2014 Apr 17;12(7):1029–1041. doi: 10.1158/1541-7786.MCR-13-0286-T

Figure 2. Generation of Cell Lines and Experimental Workflow.

Figure 2

(A) The parental RKO colon carcinoma cell line is heterozygous for the TP53 codon 72 polymorphism. Using somatic cell knockout techniques, four hemizygous cell lines were created, two having only the Pro allele, and two having only the Arg allele. A knockout cell line was also created. All six of these cell lines were (i) treated with etoposide or DMSO (control), and then subjected to RNAseq analysis and (ii) tranduced with an shRNA library, treated with nutlin-3a, and the subjected to deep DNA sequencing. (B) Western blot analysis for p53 expression levels of all six cells lines treated with either DMSO, 10 µM etoposide, 10 µM inactive nutlin-3a, or 10 µM active nutlin-3a. Note the large p53 response in all cell lines with the exception of the KO, and the slight mobility differences in Arg vs. Pro.