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. 2018 Apr 20;10(4):208. doi: 10.3390/v10040208

Figure 2.

Figure 2

Figure 2

RIG-I-N or Poly (I:C) Stimulation Directly Induces NOP53 Migration. (a) HeLa cells in 24-well plate were transfected with plasmids encoding GFP-tagged NOP53, along with poly (I:C), RIG-I, or RIG-I-N (500 ng each) for 24 h or treated with LPS at 100 ng/mL (lanes 1–5). HeLa cells were transfected with plasmids for expressing GFP-tagged NOP53 for 24 h and infected with VSV at an MOI of 5 for 10 h (lane 6). The cells were fixed with 4% paraformaldehyde. Nuclei were stained with DAPI, and NOP was visualized from the fused GFP fluorescence. Images were captured using 100× objectives with an Olympus FluoView™ FV1000; (b) Vero cells were transfected with plasmids encoding GFP-tagged NOP53 for 24 h and mock-infected or infected with PEDV or PRV at 10 MOI for 10 h. The cells were fixed with 4% paraformaldehyde. Nuclei were stained with DAPI, and NOP was visualized from the fused GFP fluorescence. Images were captured using 100× objectives with an Olympus FluoView™ FV1000; (c) HEK293T cells were co-transfected with GFP-tagged NOP53 expression plasmid and increasing doses of plasmids encoding Flag-tagged RIG-I-N. Protein levels of transfected (GFP) and endogenous (NOP) NOP53 were analyzed by immunoblotting with the indicated antibodies; (d) HeLa cells grown in 24-well plates and transfected with plasmids encoding GFP-tagged RIG-I or RIG-I-N (500 ng each) were treated with 1 μM LMB for 1 or 2 h. The cells were fixed with 4% paraformaldehyde. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI), and RIG-I or RIG-I-N was visualized from the fused GFP fluorescence. Images were captured using 40× objectives with an Olympus FluoView™ FV1000.