Figure 5. Induction of p53-bound genes by Nutlin and Wip1 inhibition.

A. Heat map of p53 at transcription start sites of genes in MCF-7 cells after Nutlin treatment. Chip data on p53-promoter-associations are displayed, red color reflecting the degree of association with p53. Group 1 indicates those genes which were significantly upregulated by both Nutlin alone and Nutlin+Wip1i treatment in the RNA-Seq analysis from Figure 4. Group 2 indicates those genes that were significantly upregulated upon Nutlin+Wip1i treatment but not by Nutlin alone. Group 3 indicates those genes that were significantly downregulated upon Nutlin and Nutlin+Wip1i and group 4 indicates those genes that were downregulated upon Nutlin+Wip1i treatment but not by Nutlin alone. B. Profiler image of p53 occupancy at transcription start sites of genes in MCF-7cells after Nutlin treatment. The profiler image (right) provides the average p53 signal obtained ± 3kb from the transcriptional start site for the genes at each of the above-mentioned conditions. ChIP-seq track data for Nutlin-3a-stimulated MCF-7 cells was obtained from p53 ChIP-Sequencing [40] and downloaded from the Gene Omibus database (ID GSE47043). C. Heat map of p53 on the TSSs of genes dependent on both p53 activity and stability. For better evaluation, we distinguished the genes from A, group 1 and 3, into two classifications, i. e. genes that were induced/downregulated further by the combined treatment in comparison to Nutlin alone (1 and 3), and the ones which were already induced/repressed by Nutlin alone to the maximum extent (2 and 4). D. Profiler image of p53 occupancy at TSSs of genes dependent on p53 activity and stability. The p53 promoter signal was aggregated along the TSSs of these genes as described in C, and a profiler image is displayed on the right.