a A general workflow developed for conducting HCC EV purification, followed by quantification of 10 HCC-specific mRNA transcripts in the purified HCC EVs. b Heatmaps depicting relative signal intensities for each gene expression of the 10 HCC-specific genes across different patient cohorts. (upper) Patients with newly diagnosed HCC (n = 46) are grouped according to Barcelona Clinic Liver Cancer (BCLC) staging system from early stages to advanced stages. (middle) Noncancer cohorts, including patients with liver cirrhosis (n = 26), chronic hepatitis (n = 25), and healthy donors (n = 23). (lower) Patients with cancers other than HCC (n = 38): cancers of nonhepatic origin metastatic to the liver (MET, n = 12); other primary cancers (n = 26), including intrahepatic cholangiocarcinoma (ICC), prostate cancer, midgut neuroendocrine tumor (NET), breast cancer, and lung cancer. Primary copy numbers are log2-transformed for each gene across all disease states. Clinical characteristics for each cohort are listed in Supplementary Tables 4–8. HBV hepatitis B virus, HCV hepatitis C virus, ALD alcoholic liver disease, NASH non-alcoholic steatohepatitis.