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Figure 2 | Veterinary Research

Figure 2

From: The activation of p38MAPK and JNK pathways in bovine herpesvirus 1 infected MDBK cells

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.

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