Fig. 3
Induction of HCC using a predefined combination of drivers. a Immunohistochemical analysis of the Fah selection marker, Gpc3, and Afp HCC markers in liver sections from Fah−/− mice treated with either control (no amiR, EGFP) or driver (amiR-mP53/1, hRasG12V) transposon constructs at 5 weeks and 5 months post-treatment. For the analysis of tumors emerging 5 months after treatment with the driver construct, a vector mixture containing 1% driver transposon vector and 99% transposon vector expressing only the Fah selection marker protein was used. Scale bars, 100 μm. b Determination of the percentage of Fah-positive hepatocytes 5 weeks after treatment by machine learning-based measurement. Data were presented as the mean ± SD (n = 3) (see Additional file 2 for individual data values and statistics). c Monitoring of endogenous p53 mRNA levels in the liver of Fah−/− mice treated with driver and control transposon constructs. Liver RNA samples were collected from Fah−/− mice at 5 weeks post-treatment and tested using a p53 mRNA-specific RT-qPCR assay. Results were normalized to measurements of the ribosomal protein L27 (Rpl27) transcript as input control and data were presented as the mean ± SD (n = 3) (see Additional file 2 for individual data values and statistics). d Monitoring the amount of transcripts A and B in the liver of Fah−/− mice treated with driver and control transposon constructs. Liver RNA samples were collected from Fah−/− mice at 5 weeks post-treatment and tested using Fah-, EGFP-, and hRasG12V-specific RT-qPCR assays. Results were normalized to measurements of the ribosomal protein L27 (Rpl27) transcript as input control and data were presented as the mean ± SD (n = 3) (see Additional file 2 for individual data values and statistics)