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. 2015 Apr 13;10(4):e0118477. doi: 10.1371/journal.pone.0118477

Fig 8. Effect of PB1-F2 stabilization on the induction of the human IFN-β promoter.

Fig 8

(A) A 293T cells were co-transfected with the human IFN-β promoter reporter plasmid pGLhIFNBpr, pRLTK (transfection efficiency normalization) and the respective PB1-F2 expressing plasmids (pPB1-F2 PR8/pPB1-F2 PR8 3KR/pPB1-F2 HK97/pPB1-F2 HK97 4KR/pPB1-F2 Stop3aa). At 24 hr p.t., the IFN-β promoter was further induced through the addition of pIC. At 15, 24 and 40 hr p.t., the supernatant was removed, and the cells were gently washed, directly lysed using passive lysis buffer (Promega) and the luciferase activity was quantified using the Dual Luciferase Reporter Assay System kit (Promega) on a BIOTEK Synergy HT luminometer. The samples were measured in triplicate and normalized to renilla luciferase activity (transfection efficiency) and firefly luciferase activity (human IFN-β promoter induction) determined for pPB1-F2 Stop3aa co-transfected samples. The data represent 3 independent experiments. The bars indicate the average value of the independent experiments, and the error bars indicate the intra-experiment SD values. The asterisk indicates a significant difference between the groups compared (* P<0.05; ** P<0.01; *** P<0.001). (B) PB1-F2 protein and Beta-actin was detected through western blotting to compare the induction of IFN-β promoter with the relative amount of the PB1-F2 in the same samples.