Figure 5

Analysis of NE treatment of reovirus virions. A. Lang virions (7.5 × 10¹⁰) were incubated with purified 25 μg/ml NE (Calbiochem) at 37°C for the indicated times (in min) and analyzed on SDS-12% polyacrylamide gels and proteins were visualized by Coomasssie staining. The mock sample (M) consisted of virions held in reaction buffer in the absence of protease for 4 h. The positions of reovirus capsid proteins are labeled. B. L929 cells were left untreated (-) or were pre-treated with 300 μM E64 and infected with Lang virions (black bars), ISVPs (white bars) or NE-treated particles (grey bars) at an MOI of 3. Viral yield was determined at 1 d p.i. C. L929 cells were left untreated (-) or were pre-treated with 20 mM NH₄Cl, 25 nM Baf or 25 μM monensin and infected with Lang virions or protease-treated particles. Cell extracts prepared at 1 d p.i. were analyzed for the expression of the viral non-structural protein μNS by immunoblotting using antiserum against μNS (diluted 1:12500).