In order to closely evaluate the immunosuppressive nature of local tumor immune microenvironment manifested in late-stage HCC patients, we transplanted fewer tumor tissues to allow the establishment of immunosuppressive microenvironment in orthotopic HCC mice as demonstrated by significantly increased numbers of immunosuppressive Tregs and elevated levels of TGF-β in blood and tumor tissues from day-21 orthotopic HCC mice compared with normal controls (Supplementary Fig. 3a). This evidence concerns the gene TGFB1 and neoplasm.