Fig. 4

Methylation levels of the 26 candidate biomarkers. A Methylation levels of the 26 candidate CpG sites in the 28 pooled cfDNA samples, with the corresponding MethylationEPIC CpG probe ID. Average methylation of the probes targeting the v2 promoter region of the SEPT9 gene (cg02884239, cg20275528, and cg12783819) is also shown. Methylation is shown as beta-values ranging from 0-unmethylated to 1-fully methylated (**differential methylation p value < 0.01; *differential methylation p value < 0.05). B Methylation levels of the 26 candidate CpG sites and SEPT9 promoter in the biomarker evaluation cohort (n = 48) of individual serum cfDNA samples. Methylation percentage was obtained through bisulfite pyrosequencing (*Wilcoxon rank-sum test p value < 0.05). C Strip-plot showing the concordance of methylation levels between pooled and individual samples. Each dot represents the methylation level of one sample. D Scatterplot shows the positive significant correlation between methylation in pooled and individual cfDNA samples for the 26 candidate CpG sites