Figure 4: AD80 induces an increased metabolism and reduced oncogenic signaling gene signature compared to sorafenib in HUH7 cells.

The AD80-induced gene signature significantly correlates to greater overall survival and lower AFP levels based on TCGA-LIHC data set. A. Heat map of metabolism and MAPK associated genes significantly upregulated or downregulated across the three groups within the indicated cell lines. Color key denotes the Z-Score (number of standard deviations from the mean) of the sample and gene row. B. Gene set enrichment analysis based on HUH7 data against the Hallmark Gene Sets, where normalized enrichment scores (NES) are shown as bars and false discovery rate (FDR) < 0.05 is marked by an asterisk. For A and B, three biological replicates per treatment near the IC₅₀ dose (sorafenib 5 μM, AD80 50 nM, DMSO 0.1%) on HUH7 or THLE5B cell lines as indicated, over a course of 24 hours, were completed. C. Kaplan-Meier survival comparison of TCGA-LIHC patient samples, separated by patients with (+) or without (−) the genes upregulated/downregulated by AD80 (Supplemental Table 3). Log-rank (Mantel-Cox) testing used to determine significance. Summary of number of patients at risk every 2.5 years, separated based on AD80 signature shown below the Kaplan-Meier. D. Box and whisker plot of TCGA-LIHC AD80 signature.