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. 2016 Nov 30;7:460. doi: 10.3389/fphar.2016.00460

FIGURE 6.

FIGURE 6

WPS inhibited influenza A/PR/8/34 (H1N1) protein expression in RAW 264.7 cells. RAW 264.7 cells were treated with WPS (100 μg/mL), interferon (IFN)-β (1000 U/mL, recombinant mouse, pre-treatment positive control), ribavirin (10 μg/mL, post-treatment positive control), or the medium only (negative control) before (A,C) or after (B,D) after viral adsorption. Influenza A virus protein levels (NS-1, PA, HA, PB-1, M1, M2) in cell lysates were analyzed by Western blotting, and β-actin expression was analyzed as an internal control. Data are presented as the mean ± SD (error bars) and are representative of three independent experiments. ∗∗∗P < 0.0005, ∗∗P < 0.005, ∗P < 0.05 compared with the mock value. RAW 264.7 cells were infected with H1N1 (multiplicity of infection = 1) before (E) or after WPS treatment (F). Cells were observed by fluorescence microscopy using an M2-specific antibody. The cells were also stained with DAPI. The merged image illustrates the cytoplasmic localization of viral M2 (red) and selected nuclei (DAPI, blue).