Figure 1. p53^R172H elevates the expression of a subset of chemokine genes.

A, Generation of Trp53^−/− isogenic cells from the parental Trp53^R172H/− cells. Trp53 gene with R172H mutation in exon-5 is deleted from intron-1 to intron-10 using CRISPR/Cas9.

B, Minus-average (MA) plot of RNA-seq transcripts showing the differentially expressed genes between Trp53^R172H/− and Trp53^−/− isogenic cells. Significantly upregulated and downregulated genes (adjusted p-value < 0.001 and four-fold change in normalized counts) are shown in blue and red, respectively. The wild-type p53-regulated genes are shown in yellow.

C, Pathways (left) and transcription factor targets (right) enriched in p53^R172H-upregulated genes. Gene Ontology analysis was performed using Enrichr¹⁰¹ against the KEGG pathway database¹⁰² (left) and TRRUST database¹⁰³ (right). Blue dots represent significant gene sets (p-value < 0.05), and the darker color represents higher significance.

D, Generation of Trp53^R172H-restored isogenic cells in Trp53^−/− cells using a Trp53^R172H cDNA expression cassette in piggyback vector (Trp53^−/− + pTrp53^R172H). An empty vector without the Trp53^R172H cDNA expression cassette (Trp53^−/− + pEV) was inserted in Trp53^−/− as a control.

E, mRNA levels of the expressed chemokine genes are shown as the z-score heatmap of RNA-seq transcripts per million (TPM) in the four isogenic cells.

F, Quantification of the three chemokine genes under p53^R172H control by ELISA in the tissue culture media of the four isogenic cells. P-values are calculated from a one-way ANOVA test followed by a post hoc test with Benjamini-Hochberg correction. Panels B, C, E, and F use Trp53^−/− clone-1 isogenic cells.