Fig. 1: Genetically-distinct murine HCCs display distinct transcriptomic and histopathological features.

a Schematic diagram of experimental design: Hydrodynamic tail vein injection (HDTVi) method used to deliver the Sleeping beauty (SB) transposon and CrispR-Cas9 constructs that enforce expression of the depicted oncogenic drivers in hepatocytes in order to generate genetically-distinct HCCs. b Kaplan–Meier survival curves of HCC-bearing mice HDTV injected with the indicated combinations of oncogenic drivers (Myc^OE/Trp53^KO n = 19, median survival of 23 days; Myc^OE/Pten^KO n = 14, median survival of 29.5 days; Nras^G12D/Pten^KO n = 15, median survival of 62 days; Nras^G12V/Pten^KO n = 14, median survival of 118 days). Nras^G12V/Pten^KO vs Nras^G12D/Pten^KO p < 0.001; Nras^G12V/Pten^KO vs Myc^OE/Trp53^KO p < 0.001; Nras^G12V/Pten^KO vs Myc^OE/Pten^KO p < 0.001; Nras^G12D/Pten^KO vs Myc^OE/Trp53^KO p < 0.001; Nras^G12D/Pten^KO vs Myc^OE/Pten^KO p < 0.001; Myc^OE/Trp53^KO vs Myc^OE/Pten^KO n.s. (non-significant). ****p < 0.0001 c Longitudinal HCC volumes of individual tumor-bearing mice (Myc^OE/Trp53^KO n = 9, Myc^OE/Pten^KO n = 6, Nras^G12D/Pten^KO n = 15, Nras^G12V/Pten^KO n = 9) determined from weekly/bi-weekly magnetic resonance imaging (MRI), and represented relative to each animal initial tumor volume. d Representative images of H&E, Masson Trichrome, and Oil Red staining performed on sectioned livers collected from end-stage HCC-bearing mice. (Scale bars = 100 µm; representative of n = 4 mice). e Principal Component Analysis (PCA) plot depicting the transcriptome differences between genetically-distinct HCC bulk tumors (Myc^OE/Trp53^KO n = 3, Myc^OE/Pten^KO n = 3, Nras^G12D/Pten^KO n = 4, Nras^G12V/Pten^KO n = 3) and control livers injected with empty vectors, hereafter referred to as control (n = 3), following RNA-seq analyses (Supplementary Data 1). f Heatmap of unsupervised hierarchical clustering depicting the geneset enrichment of the MAPK, PI3K, and MYC signaling pathways in genetically-distinct HCCs (Myc^OE/Trp53^KO n = 3, Myc^OE/Pten^KO n = 3, Nras^G12D/Pten^KO n = 4, Nras^G12V/Pten^KO n = 3) compared to control (n = 3). The color scale represents the significance of the enrichment in –log₁₀(FDR). FDR False Discovery Rate. Statistical significance was determined by log-rank (mantel-cox) test (b), two-sided hypergeometric test with multiple testing correction using Benjamini-Hochberg (f). Source data are provided as a Source Data file.