(A) mRNA levels of stemness-related genes (NANOG, OCT4, and SOX2) in human HCC cell lines of HepG2.2.15, Hep3B, PLC5 (HBV+HBsAg+), HA22T (HBV+HBsAg−), HepG2, and Huh7 (HBV−HBsAg−) with inflamed-CM treatment for 7 days (by quantitative real-time RT-PCR). The dashed line indicates gene expression in HCC cells without inflamed-CM treatment (multiple of expression = 1, control group). (B) OCT4 and NANOG levels in HCC cells with or without inflamed-CM treatment (through Western blotting). (C) Number of EGFP+ cells among OCT4 promoter-EGFP HepG2 and HepG2.2.15 cells with and without inflamed-CM treatment. EGFP+ HepG2.2.15 cells show mesenchymal-like cell morphology. EGFP, enhanced green fluorescence protein; Ph, phase image. (D) Number of EGFP+ cells (per 103 cells) among HepG2 and HepG2.2.15 cells with and without inflamed-CM treatment. (E) The EGFP level in HepG2.2.15 cells with inflamed-CM treatment (by Western blotting). (F) The relative luciferase activity of OCT4 promoter-luciferase HepG2.2.15 cells with inflamed-CM treatment. *P < .05, **P < .01, ***P < .001, by t-test.