Fig. 3

Effects of EF on the proliferation of HFLS-RA cells with and without TNF-α stimulation. A Toxicity of EF in cultured cells was tested at 24 and 48 h. B Anti-proliferative effects of EF against TNF-α-stimulated HFLS-RA cells exhibited a concentration-dependent relationship. HFLS-RA cells were incubated with TNF-α (10 ng/mL) and EF (25, 50, 100, 200, 400, 800, 1600 µg/mL) for 24 h. The CCK-8 assay was employed to determine the extent to which cell proliferation was inhibited (%). Data are the mean ± SD (n = 4), *p < 0.05, **p < 0.01, ***p < 0.001, vs. TNF-α cells (HFLS-RA cells treated by TNF-α alone). ^#p < 0.05, ^##p < 0.01, ^###p < 0.001, vs. control cells. C A cell-colony assay was used to investigate the effect of EF on the proliferation of HFLS-RA cells. D Data are the mean ± SD (n = 4), *p < 0.05, **p < 0.01, ***p < 0.001, vs. control cells. E HFLS-RA cells were seeded onto glass coverslips, serum-starved, and then stimulated for 24 h with TNF-α (10 ng/mL) or EF (400, 800, 1600 µg/mL). Then, cells were stained with the EdU Cell Proliferation Kit. Hoechst (blue) and Azide594 (red) were visualized by fluorescence microscopy (×200)