FIG 1.

JUNV C1-mediated activation of the NF-κB and Erk1/2 pathways is dependent on TLR2 signaling. (A) Primary macrophages with deletions in TLR genes were infected with JUNV C1 or treated with LPS, LPS plus polymyxin B (Pmx), or PAM, and at 2 hpi, protein lysates were prepared and analyzed by Western blotting, using antibodies against total and phosphorylated (p-Ser486) NF-κB (p65). (B) Primary macrophages were treated with UO126 for 1 h, followed by infection with JUNV C1 or LPS. At 2 hpi, cell extracts were analyzed for phosphorylated Erk1/2 by Western blotting. (C and D) At 2 hpi, RNA was isolated from cells treated as indicated, and IFN-β (C) and TNF-α (D) RNA levels were quantified by RT-qPCR. (E) Supernatants from the same cells were used to quantify TNF-α levels by ELISA. The bar graphs represent the averages and standard deviations from three technical replicates. These experiments were performed twice, with similar results.