Fig. 1.

Transcriptional regulation and protein expression of chicken TLR3, TLR7, MDA5, IFNβ, MyD88, TRIF, and NF-κB in IBV-infected CEK cells and SPF chickens. (A-E) CEK cells were inoculated with IBV strain AH, TM, or H52 at an MOI of 1. The cells were collected at 12, 24, 36, or 48 hpi. The expression of chicken TLR3, TLR7, MDA5, IFNβ, and IL-6 mRNA was detected using qPCR. (F) The expression of IFNβ protein in cell culture supernatants was detected using an ELISA assay. (G-I) The expression of MyD88, TRIF, NF-κB, and β-actin proteins was detected using anti-MyD88, anti-TRIF, anti-NF-κB, and anti-β-actin antibody. (J-M) SPF chickens were challenged with a 0.2-ml suspension containing 10⁶ EID₅₀ of IBV, and the kidneys of these chickens were collected at 5 dpi. The expression of chicken TLR3, TLR7, MDA5, and IFNβ mRNA was detected using qPCR. Data were analyzed using unpaired t-tests or ANOVA, followed by Dunnett’s multiple comparison test, using GraphPad Prism (GraphPad Software, San Diego, CA). The significance of the differences between the infected and mock-infected groups is indicated by the p-value