Fig. 2

High-performance liquid chromatography (HPLC) analysis using the newly developed anion-exchange column for specimens of normal liver tissue (NLT) (n = 30) and non-cancerous liver tissue showing non-alcoholic steatohepatitis (NASH) derived from partial hepatectomy specimens from patients with hepatocellular carcinoma (HCC) (NASH-W) (n = 20) in the initial cohort. A Chromatograms of PCR products encompassing the marker CpG sites (cg09580822, g15050398, cg18210511, cg09580859 and cg13719443) for representative tissue samples (black). Gene names are shown on the left side. N/A: not annotated (the probe was located within the intergenic region). Chromatograms of fully unmethylated and fully methylated control DNAs are shown in blue and red, respectively. For cg09580822 and cg15050398, chromatograms with a “single peak” were obtained in both NLT and NASH-W. For cg18210511 and cg09580859, NLT showed a “bimodal peak” pattern, whereas a “single peak” pattern was observed in NASH-W. For cg15050398, NLT showed a “single peak with a shoulder” (indicated by an arrow) pattern. B Scattergrams of the relative DNA methylation rates obtained by HPLC analysis for cg09580822, g15050398, cg18210511, cg09580859 and cg13719443. Such rates are described in “Results” section and differed significantly between NLT (black) and NASH-W (red) samples. P values were obtained by Welch’s t test