Figure 2

INA binds to the viral RNA and renders it non-infectious in-vitro . (A) L-cells were transfected with RNA isolated from infectious EMCV (E-RNA) or EI₁₀₀i (EI₁₀₀i-RNA) or mock transfected. EMCV-3D gene specific RT-PCR was done on the total RNA isolated from cells after 48 h post transfection to test for EMCV replication. (B) Virus replication in cell supernatants was measured by standard plaque assay. No virus replication was detected in the supernatant of the cells transfected with the EI₁₀₀i-RNA. Virus titer in cells transfected with E-RNA was similar to the samples infected with control live EMCV virus. C) RNA was isolated from EMCV (E-RNA), EMCV treated with 100 μM I125-labeled INA (EI₁₀₀-RNA), and EMCV treated with 100 μM I125-labeled INA in combination with UV (EI₁₀₀i-RNA). Co-localization of INA with RNA was tested by overlapping silver staining of RNA (upper panel) with autoradiograph of I125-labeled INA (lower panel). D) RNA isolated from EI₁₀₀i (EI₁₀₀i-RNA) was treated with RNase t1 or Proteinase K to test the binding of INA to EMCV RNA.