AMPK restricts RVFV RNA replication.

<p><b>A.</b> Northern blot of genomic S segment and N mRNA from RVFV (MOI 1) grown in WT or AMPKα1/AMPKα2^−/− MEFs for 4, 8, or 12 hours. A representative of triplicate experiments is shown. <b>B–C.</b> Quantification of RVFV mRNA (<b>B</b>) or genomic RNA (<b>C</b>) in WT or AMPKα1/AMPKα2^−/− MEFs displayed as the normalized fold change from WT 4 hours. A representative of triplicate experiments is shown. <b>D.</b> RVFV binding assay. RVFV (MOI 10) was bound to WT or AMPKα1/AMPKα2^−/− MEFs at 4°C for 1 hour, then washed, and treated with PBS or trypsin to remove bound virus. qRTPCR was performed on isolated RNA to detect RVFV S genome. Data are displayed as the average ΔΔCT of triplicate experiments normalized to GAPDH control. * indicates p<0.05. <b>E.</b> 2DG (12 mM), A769662 (100 uM) or Ammonium Chloride (NH₄Cl, 12 mM) was added either 1 hour prior to infection with RVFV (MOI 1), with infection, or 1, 2, or 4 hours post infection. After 10 hours of infection cells were fixed and processed for immunofluorescence. Data are displayed as the average percent infection relative to the post entry level of infection (NH₄Cl added at 4 hpi) ± SD from triplicate experiments. * indicates p<0.05.