Figure 5.

Rg1 inhibits the NF-κB pathway activated by PRRSV infection. (A) The expression and phosphorylation level of proteins involved in NF-κB pathway are analyzed in uninfected (Mock) and PRRSV XH-GD (0.1 MOI) infected Marc-145 cells treated with or without Rg1 (200 μM), samples were collected at 24 h.p.i. As a positive control, cells were cultured in DMEM and supplemented with LPS (2.5 μg/mL) at 37 °C for 6 h, and then the medium was changed to medium containing 0 or 200 μM Rg1 for 18 h. The western blotting data of each target protein represents three independent experiments with similar results. (B) Marc-145 cells were grown on glass cover slips and cultured in medium at 37 °C for 24 h, and then infected with PRRSV XH-GD (0.1 MOI). After virus infection, cells were incubated in fresh DMEM supplemented with or without 200 μM Rg1 for 24 h. Cells were washed twice with PBS and performed immunostaining by using anti-P65 antibody and red-fluorescent Alexa Fluor 594-conjugated goat anti-mouse IgG antibody. Nuclei were counterstained with DAPI. P65 protein deposited in the nucleus was indicated by yellow arrow. (C) Schematic model of Rg1 affect NF-κB signaling pathway upon PRRSV infection.