Figure 4.

Differential expression of the TLR downstream genes in response to West Nile virus. (A) TLR-associated genes mRNA expression in fold change. The ΔΔCt (ΔΔCt = ΔCt_WNV − ΔCt_mock) values were calculated by subtracting the ΔCt of genes in mock-inoculated PBMCs (n = 3). The bar graph showed the expression of genes in WNV-infected PBMCs over mock-inoculated PBMCs (fold change: the normalized expression value of a gene in WNV-stimulated cells/the normalized expression value of a gene in mock-inoculated cells). Bars without common superscripts (a,b) denote statistical difference among time points (p < 0.05). (B–E) Relative expression of TLRs-associated genes mRNA, accounting for the effects of culture conditions on gene transcription in WNV- and mock-inoculated rabbit PBMCs (n = 3). To compare the normalized expression of TLR-associated genes from PBMCs harvested at each time point to their respective expression levels before either WNV inoculation or mock inoculation, the ΔΔCt values were calculated by subtracting ΔCt of genes in fresh-isolated PBMCs from the ΔCt of genes in WNV- or mock-inoculated PBMCs at each time-point (for WNV-stimulated PBMCs, ΔΔCt_WNV = ΔCt_WNV − ΔCt_fresh; and for mock-inoculated PBMCs, ΔΔCt_mock = ΔCt_mock − ΔCt_fresh). A time-dependent relative expression patterns of (B) TRAF3, (C) STAT1, (D) IRF7, and (E)IRF9 mRNA in WNV-challenged rabbit PBMCs at different time points. Line graphs without common superscript differ significantly (p < 0.05). Upper case letter denotes difference of a gene expression among the time points in WNV-challenged cells; lower case letter denotes difference of a gene expression among the time points in mock-inoculated cells. *indicates the difference of a gene expression between WNV- and mock-challenged cells in the same time point (p < 0.05).