Specific inhibition of phosphorylation and activation of swine MAP kinases and NFκB in H1N1pdm-infected 3D/3 cells.

<p>Swine macrophages were pretreated with U0126, SB230058, and InSolution JNK Inhibitor, which are inhibitors of ERK1/2, p38, and JNK1/2, respectively. The final concentrations of the inhibitors were 10 µM, 5 µM, and 50 µM for U0126, SB230058, and the InSolution JNK Inhibitor, respectively. After 1 hr incubation, the treated cells were infected with H1N1pdm at an MOI of 1, and the cell lysates were prepared at 18 and 30 hrs post infection. The cell lysates were subjected to SDS-PAGE and western blot analysis with anti-phospho-ERK1/2 and anti-phospho-JNK1/2 antibodies. <b>A.</b> Specific inhibition of ERK1/2 phosphorylation and activation by U0126. <b>B.</b> Specific inhibition of JNK1/2 phosphorylation and activation by InSolution JNK inhibitor. <b>C.</b> Phosphorylation and inhibition of NFkB activation in infected 3D/4 cells. The cells were pre-treated with the inhibitors 1 hr prior to infection. The final concentrations for the NFκB and IKK inhibitors were 10 nM and 10 µM, respectively.