Figure 5

The mabJ2 dsRNA replication assay is compatible with high content image based siRNA infection screens. (A) Overview montage of an example siRNA screening plate. HeLa cells were infected with HRV14 and stained with a rabbit polyclonal antibody (rpc, green) raised against purified viral capsid, mabJ2 recognizing dsRNA (red) and nuclei (DAPI, blue). One out of nine images per well is shown for each siRNA, which are not specified here. (B) Examples close-ups from wells treated with HRV-targeting (HRV siRNA), no siRNA, or scrambled siRNA, followed by staining as described in (A). Merged colors are shown above, single channel micrographs are in black and white. Scale bars 100 μm. (C) Normalized HRV16 infection index (log2 transformed) determined by automated microscopy/analysis from three independent siRNA screens. Infection was measured either by mabR16-7 recognizing a VP2 epitope or mabJ2 recognizing replicated dsRNA.