Fig. 1.

The effect of Huaier and 5-FU treated alone or in Combination on cell proliferation and colony formation in Huh28 cells. a and b After being treated with Huaier (1.5-24 mg/ml) or 5-FU alone (7.5-120 μg/ml) in different concentrations for 24 h and 48 h, the cell viability was measured with CCK-8 assay. c and d Cell proliferation ability was also assessed after cells were treated with Huaier and 5-FU alone or in combination for 24 h and 48 h. e and f The inhibiting effects on colony formation by treated monotherapy or in combination were shown, and the colony numbers were calculated. g and h The protein expressions of Ki67 and PCNA in Huh28 cells after 48 h treatment with Huaier and 5-FU alone or in combination were measured with western blot, and relative optical density of the proteins expression was analyzed using Image J. All data are expressed as the mean ± S.D. of three separate experiments. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, compared with control group, while ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, compared with combination group. i The panel labels “24h” and “48” represent the samples were detected after treatment 24 hours and 48 hours respectively. The panel labels “Control”, “Huaier”, “5-FU” and “Huaier+5-FU” represent the samples were treated by neither of two drugs, Huaier extract, 5-Fluorouracil and combination of two drugs respectively