Figure 5.

A528R inhibits the antiviral and antibacterial effects of TLR8-NF-κB signaling. (A–B) PAMs grown on glass coverslips in 12-well plate (3 × 10⁵ cells/well) were transfected with A528R-HA (1 µg), A528RΔ54-83 (1 µg), or empty vector. At 24 h post transfection, cells were stimulated with R848 (5 µg/mL) for 12 h. The cells were infected with VSV-GFP at the MOI 0.01 for 12 h. The VSV replicative GFP signals were observed by fluorescence microscopy (A) and the GFP expression was analyzed by Western blotting. The GFP gray density were quantified, and the values were shown under the respective GFP bands after normalization of β-action (B). (C–D) PAMs grown in 12-well plate (3 × 10⁵ cells/well) were transfected and treated with R848, as in A–B. The cells were infected with Staphylococcus aureus at MOI of 15. At 6 h post infection, the cells were harvested and lysed. The diluted lysates were spread on LB agar plate. The representative picture of bacterial colonies on overnight grown plates were shown (C), and the colony numbers of each sample were counted and shown as the chart form (D).