poSn-DAP12 pathway mediates inhibition of NF-κB in response to PRRSV.A, B PAMs were pre-incubated with PRRSV (MOI = 1) for 1 h. After washing with serum free medium extensively, the cells were transfected with the indicated sipoSns for 35 h. poSn knockdown efficiencies were examined by RT-qPCR (A). IB analysis was performed to examine the expression of PRRSV N protein, and the activation of IRF-3 and NF-κB (B). C, DDAP12 knockdown PAMs were infected with PRRSV (MOI = 1) for indicated time periods (0, 3, 6, 9, 12, 24 h). IB was conducted to determine the abundance of PRRSV N protein (C ,D), and the activation of NF-κB (C) and IRF-3 (D). E DAP12-overexpressed CRL-2843-CD163 cells were infected with PRRSV (MOI = 5) for the indicated time periods (0, 3, 6 h). IB was adopted to detect IκB-α degradation and phosphorylation of IRF-3 and p65. IB panels were representative from three independent experiments. RT-qPCR data were indicated as mean ± SEM. Statistical significances were shown by the Student t test: ***p <0.001.