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Figure 3: Hepatoma cell lines MHCC97H and Hep3B treated with SYY (2 mg/mL) for 4 weeks exhibited decreased stemness. (a) Tumor cells that were treated with SYY for 4 weeks showed reduced expression of CSC-related markers compared with their parental cell lines by flow cytometry. (b) RT-PCR and Western blot analyses confirmed the decreased expression of CSC-related markers in SYY-treated cell lines. (c) The transwell assay demonstrated that MHCC97H-SYY and Hep3B-SYY cells migrated through the basement membrane less efficiently than the parental cell lines MHCC97H and Hep3B. (d) Colony formation assay demonstrated that MHCC97H-SYY and Hep3B-SYY had a significantly lower proliferation rate and colony-forming ability than untreated parental cells. (e) MHCC97H-SYY cell lines also showed diminished subcutaneous tumor growth capacity and pulmonary metastasis compared with parental cells in nude mouse models.