Mouse mab j2

HeLa cells were grown on coverslips overnight and infected with NDV strain Herts/33 at an MOI of 1 or stimulated with poly (I:C) for 12 h. Cells were fixed in 3% paraformaldehyde, permeabilized with 0.5% Triton X-100 for 10 min, incubated in blocking buffer (3% BSA/PBS), and then stained with mouse mAb J2 (Scicons, Hungary) followed by Cy3-labeled goat anti-mouse IgG (Beyotime). Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI) (Thermo Fisher Scientific Inc., Rockford, IL, USA) at a dilution of 1:500 for 10 min, and then the coverslips were mounted on slide glasses and visualized using a fluorescence microscope (Nikon Eclipse 80i; Nikon, Tokyo, Japan).

The following reagents were purchased: mouse mAb J2 (Scicons); Alexa Fluor 488-goat anti-mouse IgG secondary antibody (Molecular Probes, Life Technologies); Phi6 dsRNA (Finnzymes, Thermo Fisher Scientific); dsRNA ladder (New England Biolabs); High MW poly (A:U) and poly (I:C) (InvivoGen); GeneRuler 1 kb DNA Ladder (Thermo Fisher Scientific); Silencer negative control 1 siRNA (Ambion, Life Technologies); Strep-Tactin conjugated to horseradish peroxidase (IBA Lifesciences); pNPP (Sigma-Aldrich), mMESSAGE mMACHINE T7 transcription kit (Ambion, Life Technologies), Lumi-Light western blotting substrate (Roche Diagnostics).