Fig. 6: Transcriptomic analysis reveals CP-673451 may affect KRAS/NF-κB signalling mechanism in U87 GBM cells.

A Hierarchical clustering analysis was performed using DESeq2. Dark to light colour code refers to the distance metric used for clustering. Dark blue represents the maximum correlation. B Volcano plot illustrating the Log2 fold change of genes that are altered upon CP-673451 treatment of U87 cells. The Log10 of p value, for significance in fold change, is plotted on the y-axis. Points on the plot refer to genes and are coloured according to significance. Red are genes significant by both p value and log2 fc, blue are significant only by p value but not log2 fc, green are significant only by log2 fc, grey are NS. C qRT-PCR performed on U87 cells treated with CP-673451 for 48 h validating genes identified from RNAseq as being upregulated (n = 5) and downregulated (n = 5). All genes show significance compared to control levels. D GSEA was performed to determine whether treatment of U87 cells with CP-673451 altered key Hallmark pathways curated by mSigDB. Genes predicting pathway activation of the KRAS and TNFA-NFκB signalling were significantly upregulated in U87 cells treated with CP-673451 in comparison to DMSO (FDR ≤ 0.1). E Heatmap from GSEA analysis illustrating expression of genes within the leading edge of the KRAS and TNFA-NF-κB Hallmark signatures. The mean ± SD of n = 3 independent experiments is shown *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001 (two-tailed t-test).