We cultured five hepatoma cell lines (Hep3B, HepG2, LM3, Huh7, and MHCC97H) and normal liver cells LO2, extracted RNA and protein from the cells, and detected the expression of TM4SF1 at the RNA and protein levels using qPCR and Western blot (WB), respectively (Fig. 1E and F). This evidence concerns the gene TM4SF1 and hepatocellular carcinoma.