Phenotypic characterization and inflammation in c-Jun~Fra-2hep livers.  A. Principal component analysis (PCA) of RNAseq data. PC1 and PC2 account for 62.5% of sample variability. Individual samples from 2 and 9 months of transgene expression are depicted with circles and squares, respectively and the correspondence between non-tumoral (NT) and tumoral samples from the same mouse indicated with dotted arrows.  B. Normalized enrichment scores in c-Jun~Fra-2hep liver tumors relative to controls (RNAseq, n=3 controls and 4 mutants) compared with human and mouse liver cancer signatures by gene set enrichment analysis (GSEA). False discovery rate (FDR) q-val <0.0029 for upregulated (red) and downregulated (green) genes. Data are ordered by FDR in each comparison. C. Ki67 and Cyclin D1 IHC in c-Jun~Fra-2hep liver sections. Bar = 200 mm. D. IHC quantification of Ki67 in hepatocytes (hep) and immune cells (immune) and Cyclin D1 in hepatocytes in liver sections of c-Jun~Fra-2hep mice and controls at 2 months of transgene expression. E. F4/80 (macrophages), Pax5 (B cells) and CD3 (T cells) IHC in liver sections of c-Jun~Fra-2hep mice and controls. Bar = 100 mm, tumors (T) are indicated by a dotted line and arrows point to positive nuclei. F. qRT-PCR quantification of immune-cell markers in c-Jun~Fra-2hep livers at 2 months of transgene expression compared to controls. G. CIBERSORTx deconvolution of liver myeloid cells in c-Jun~Fra-2hep liver RNAseq datasets at 2 (control: C, n=6, mutant: NT, n=3) and 9 (control: C, n=3, mutant: NT, n=3, T: tumors, n=4) months. H. Top enriched immune-related MSigDB Hallmark signatures in mutant groups compared to their respective controls (RNAseq, 2 months: n=6, 9 months: n=3) by GSEA. Normalized enrichment scores (NES) are shown as bars and FDR q-values (–log10) as dot plots. Data are ordered by NES in the 2 months dataset. I. Immunoblot for total and phosphorylated Stat3 and NF-κB p65 in liver extracts from c-Jun~Fra-2hep mice (non-tumoral and tumors) and controls. Tubulin is used to control loading. Bars = means ±SEM. * p<0.05, ** p<0.01, *** p<0.001 (t-test to controls).