Figure 2

The JNK but not p38MAPK signaling related to BHV-1 replication. A, B JNK inhibitor inhibited BHV-1 replication. MDBK cells were first pretreated with inhibitor SP600125 A specific for JNK or inhibitor SB203580 B specific for p38MAPK at various concentrations for 1 h, and then infected with BHV-1 at an MOI of 1. At 24 hpi, virus yield was titrated using the TCID₅₀ assay. C Screening siRNA target for JNK1 and JNK2. MDBK cells in 6-well plates were transfected with the indicated siRNA of 100 pM using transfection reagent siRNA-Mate (Genepharma) according to the specifications. At 48 h post transfection, the cells were lysed and subjected to Western blotting with antibody against JNK. D Knock down of the expression of JNK1 with siRNA1 moderately inhibited BHV-1 replication. MDBK cells in 24-well plates were transfected with siRNA1, siRNA3 and control siRNA of 20 pM using transfection reagent siRNA-Mate. At 48 h post transfection, the cells were infected with BHV-1 at MOI of 1 for 1 h, after extensive washing with PBS, fresh medium was replaced for further incubation of 24 h. Virus yield was tittered with TCID₅₀. The data are from three independent experiments. Statistical analyses were performed using the Student’s t test (P < 0.05 vs. control). E Knockdown JNK1 by siRNA1 reduced JNK1 phosphorylation induced by BHV-1 infection. MDBK cells in 6-well plates were transfected with siRNA1 and control siRNA, respectively. At 48 h post-transfection, the cells were infected with BHV-1 at MOI of 10 for 0.5 h. The cell lysates were prepared for Western blotting analysis. Data are representative of two repeats.