Avian infectious bronchitis virus disrupts the melanoma differentiation associated gene 5 (MDA5) signaling pathway by cleavage of the adaptor protein MAVS

Yu, Liping; Zhang, Xiaorong; Wu, Tianqi; Su, Jin; Wang, Yuyang; Wang, Yuexin; Ruan, Baoyang; Niu, Xiaosai; Wu, Yantao

To further understand the role of chMDA5 and chMAVS during IBV infection, chMDA5 and chMAVS were knocked down in CEK cells, then the cells were infected with IBV at an MOI of 1. IBV in supernatants Fig. 4 Dose-dependent antiviral cytokine potency of IBV. CEK cells were inoculated with 10 −2 , 10 −1 , 10 0 , 10 1 and 10 2 MOI of IBV, then the cells were harvested at 36 h post-treatment for RNA extraction. The IBV genome load (a) was quantified by RT-qPCR. The mRNA levels of chMDA5 (b), chIFN-β (c), chIFN-λ (d) and chMx (e) were evaluated by RT-qPCR. All gene expression was calculated as the fold change relative to uninfected control cells and normalized against a housekeeping gene (β-actin). Data are presented as the mean ± SD (n = 3, 3 wells on same plate) (* P ≤ 0.05; ** P ≤ 0.01) was titrated onto CEK cells at 48 h post-infection while the IBV genome loads were quantitated by real-time PCR. The results indicated that the observed viral replication in chMDA5 and chMAVS knockdown cells was the same as in the control group (Fig. 7b and c) . Overall, inhibition of chMDA5 and chMAVS expression has no effect on the replication of IBV.