BoHV-1 infection of MDBK cells induced Akt phosphorylation via a PI3K-dependent pathway. (a) Time course for BoHV-1 stimulation and Akt phosphorylation. Growth arrested cells were infected with BoHV-1 and processed for Western blotting at the pi time indicated. Mock infected cells were used as a control. (b) Time course for BoHV-1 stimulation and Akt phosphorylation at the early time. (c) PI3K specific inhibitor Ly294002 suppressed Akt phosphorylation induced by the virus infection. Cells preincubated with medium in the absence of Ly294002 and then mock infected or infected with the virus were used as the negative and positive control, respectively. Growth arrested MDBK cells were pretreated with Ly294002 and infected with BoHV-1 at an MOI of 2 in the presence of the inhibitor, and then processed for Western blotting at the pi time indicated with phosphor-Akt antibody. Western blotting was performed with Akt antibody to detect the protein loading. Each experiment was repeated three times, and representative results are shown.