Cross-talk between MAP kinases and NFκB pathways in H1N1pdm-infected swine macrophages.

<p><b>A.</b> Inhibition of NFκB activation in H1N1pdm-infected 3D/4 cells. The cells were pretreated with U0126, SB230058, and InSolution JNK Inhibitor, 1 hr before H1N1pdm infection. Cell lysates were prepared at 12 and 24 hrs post infection and subjected to SDS-PAGE and western blot analysis with anti-phospho-NFκB (p65) antibody. <b>B.</b> Induction of RIG-1 and MDA-5 in infected swine macrophages. Total RNA was prepared from infected cells for reverse transcription. cDNA was used for realtime PCR with RIG-1 and MDA-5 primers. <b>C–D.</b> Regulation of RIG-1 and MDA-5 induction by MAP kinases. The cells were pretreated with U0126, SB230058, and InSolution JNK inhibitor, 1 hr before H1N1pdm infection. Total RNA was prepared from infected cells for reverse transcription. cDNA was used for realtime PCR with RIG-1 and MDA-5 primers to measure fold changes of RIG-1 and MDA-5 transcripts in treated swine macrophages. Each assay was repeated at least twice. Data show mean fold change plus standard deviation of two or three independent assays. *p<0.05, Student's t-test.