Fig. 1

Cancer stem cell (CSC) properties of sphere cells in HCC cell lines. a Survival rates of Huh7 (left) and Hep3B (right) after 80 μM 5-FU (upper), 5 μM Sorafenib (middle), or 2 μM Doxorubicin (lower) treatment were evaluated by CCK8 assay. b Representative photographs of the plates containing colonies derived from 2000 sphere or parental normal Huh7 (upper) and Hep3B (lower) cells. Colony formation experiments were performed in triplicate (mean ± SD). c Representative NOD/SCID mice with subcutaneous tumors from sphere Huh7 cells and H&E staining of subcutaneous nodules. Scale bar 1 cm. d Expression levels of EpCAM, CD133, ATP-binding cassette sub-family G member 2 (ABCG2) and CD90 among the 1st, 2nd, 3rd sphere and differentiated sphere cells in Huh7 (left) and Hep3B (right) cells. Results were normalized according to the expression of parental cells. All experiments were done in triplicate. e Evaluation of sphere formation rates in three sequential generations of Huh7 and Hep3B cells. f Expression of epithelial cell adhesion molecule (EpCAM) and CD133 in 2nd sphere and parental normal Huh7 (left) and Hep3B (right) cells. Scale bar 100 μm