Fig. 2

Genome-wide siRNA screen results. a Primary siRNA screen results using the minigenome system. A classical minigenome assay in the presence of host factor-directed siRNAs targeting each gene in the human genome in triplicate was performed in HEK293 cells. Reporter activity relative to the negative siRNA control is shown on a log scale for both nano-luciferase (nluc; reflects minigenome replication and transcription) and Firefly luciferase (FF; reflects control plasmid-driven gene expression). Results in the presence of siRNAs targeting the host factors CAD, DDX39B, and NXF1, which showed a high likelihood to be true positive hits in a follow-up analysis, and ZAP, which has been previously shown to be an antiviral factor, are highlighted as colored shapes, as indicated. b Secondary assay results based on infection with a luciferase-expressing EBOV. Cells were infected with a recombinant EBOV expressing Firefly luciferase (FF) in the presence of selected host factor-directed siRNAs from the primary screen shown in a). In parallel, an ATP-based cell viability assay was performed using UltraGlo luciferase activity as a readout. Mean results from 4 biological replicates from 2 independent experiments are shown relative to the negative control siRNA (neg. siRNA). Results in the presence of siRNAs targeting the host factors CAD, DDX39B, and NXF1 are highlighted as colored shapes, and controls (mock: no infection; no siRNA; anti-L: siRNA directed against EBOV L; killer siRNA: cytotoxic siRNA as a transfection control) are shown as empty shapes, as indicated