FIG 2.

Upregulation of RIG-I and MDA5 expression by JUNV C1 is TLR2 dependent. RIG-I and MDA5 levels were analyzed by Western blotting of extracts from primary macrophages infected with JUNV C1 or treated with LPS, LPS plus polymyxin, or PAM. (A) Primary macrophages derived from wild-type mice were infected with JUNV C1, and at 2 hpi, protein lysates were analyzed by Western blotting, using antibodies against RIG-I and MDA5. (B) JUNV C1-infected macrophages derived from wild-type, TLR2^−/−, and TLR4^−/− mice were harvested at 24 hpi, and the proteins were analyzed by Western blotting with antibodies against RIG-I and MDA5. A cellular protein that is detected nonspecifically by the MDA5 antibody was used as a loading control. Shown below the lanes are the relative levels of protein in each sample relative to levels in mock-treated cells. Quantification of RIG-I and MDA5 levels was done by using ImageJ software. These experiments were done twice, with similar results.