Biphasic activation of both JNK and p38MAPK pathway in BHV-1 infected MDBK cells. A Time course of altered phospho-JNK(Thr183/Tyr185) and phosphor-p38MAPK(Thr180/Tyr182) following BHV-1 infection. Growth arrested MDBK cells were mock infected or infected with BHV-1 at an MOI of 10. At the indicated time points, cells were lysed and subjected to Western blotting analysis using the indicated antibodies. B UV-irradiation inactivated virus also induced activation of both JNK and p38MAPK. MDBK cells were infected with BHV-1 (MOI = 10) or with equal amounts of UV-irradiation inactivated virus for 30 min. The cells were lysed for Western blotting. C Uninfected cell lysates could not stimulate p38MAPK and JNK signaling. To get uninfected cell lysates and crude virus, MDBK cells were mock infected or infected with BHV-1 (MOI = 1) without serum in the medium. At 48 hpi, the cell cultures were subjected to frozen-thawing and centrifugation at 15 000 rpm for 10 min. The supernatants were collected and used for the treatment of MDBK cells. At 0.5 h post-treatment, the cells were lysed for Western blotting analysis. Data are representative results of three independent experiments.