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. 2020 Apr 30;11:678. doi: 10.3389/fimmu.2020.00678

FIGURE 5.

FIGURE 5

cMDA5 is an indispensable sensor of RNA ligands in chicken cells, whereas cTLR3 serves only a complementary function. (A) cMDA5 knockout (KO), cTLR3 KO, and MDA5/TLR3 double knockout (DKO) result in a decreased IFN-β promoter activity in response to poly I:C. Wild type (WT), cMDA5 KO, cTLR3 KO, and MDA5/TLR3 DKO DF-1 cells were co-transfected with the cIFN-β-Luc vector (200 ng) and pGL4.53 (20 ng) and then stimulated with poly I:C for 24 h. After stimulation, the cells were harvested for use in a luciferase reporter assay. (B) WT, cMDA5 KO, cTLR3 KO, and MDA5/TLR3 DKO DF-1 cells were stimulated with poly I:C for 24 h. After stimulation, the cells were harvested and the relative expressions of mRNAs encoding interferon-stimulated genes Mx1, IFITM3, OASL, and PKR were assessed by RT-qPCR. The data are represented as the mean ± standard deviation (n = 3). Data were analyzed using two-way ANOVA with Bonferroni’s multiple comparison (A,B). *P < 0.05, **P < 0.01, and ***P < 0.001. ns, non-significant.