FIG. 4.

Examination of the relationship between ODN2006 and poly(I-C) and their effects on TLR3 signaling. (A) Effects of the order of addition of poly(I-C) and ODN2006 on luciferase activity in transfected HEK 293T cells. The times of addition are shown at the bottom of the graph. (B) Comparison of the route of introduction and the inhibitory effects of ODNs. The ligands were either added to the cell culture medium (m-) or transfected into the cells by using Lipofectamine (t-), as indicated. The means of the results are shown above the bars, and the error bars show standard errors of the means. (C) The secretion of 3ECD is not affected by the transfection of ODN2006 or dODN into HEK cells. Where ODN2006 was used in the absence of Lipofectamine (LF2K), it was added to the cell culture medium. “V” indicates a sample transfected with the empty vector. The upper band is the glycosylated 3ECD that was secreted into the medium. The fainter lower band was found to be inside the cell (S. Hoose, data not shown). pIC, poly(I-C); 2006, ODN2006; +, present; −, absent.