Figure 2

Characterization of CD133⁺ Huh7 and CD133^- Huh7 cells. (A) Re-analysis of FACS-sorted CD133⁺ and CD133^- Huh7 cells. (B) RT-PCR analysis of CD133⁺ and CD133^- Huh7 cells. (C) Immunofluorescent staining of CD133⁺ and CD133^- Huh7 cells with anti-β-catenin and anti-CK19 antibodies. (D) Formation the anchorage-independent spheres in stem cell medium. (E) Quantitative analysis of sphere-forming efficiency of CD133⁺ and CD133^- Huh7 cells. (F) Immunofluorescent staining of CD133⁺ and CD133^- Huh7 cells with anti-β-catenin antibody. Arrowed cells showed nuclear staining of β-catenin. (G) Huh7 cells were transfected with equal amounts of either Super8xTOPFlash reporter or Super8xFOPFlash reporter constructs. After 48 hours, the TopFlash/FopFlash luciferase activity of CD133⁺ and CD133^- Huh7 cells was measured. Values shown are relative luminescence. (H) Huh7 cells were treated with different concentration of 5-fluoroucacil (5-FU) or Sorafenib for 48 hours. The cell survival was determined using MTT assay. (I) Huh7 cells were treated with 160 μM of 5-FU or 3 μM of Sorafenib for 48 hours. The expression of CD133 in survived cells was analyzed by flow cytometry.