The impact of E2-p7 processing on the interaction of NS2 with NS3 and E2. (A) The interaction of NS2 with NS3 and E2 was determined by Western blot analysis following NS2 pulldown assays. (B) The level of immunoprecipitated NS2 (IP-NS2) was normalized to that of NS2 present in input lysates (NS2-input). IP-NS2 (normalized with by NS2-input) from HJ3-5 was set to 100. The standard deviations from five different experiments are shown. Asterisks indicate statistically significant differences between a and b, c, d, or e paired values: ***, P < 0.0005; **, P < 0.005; *, P < 0.05. The differences with a P value of >0.05 were considered not significant (ns). (C) The same experiment as shown in panel B except that the level of immunoprecipitated NS3 (IP-NS3) was normalized to that of NS3 present in input lysates (NS3-input). Results from three different experiments are shown. (D) The data shown in panel C were normalized to those shown in panel B from three different experiments to determine the efficiency of NS2 and NS3 interaction. (E) The same experiment as shown in panel B except that the level of immunoprecipitated E2 (IP-E2) was normalized to that of E2 present in input lysates (E2-input). (F) The data shown in panel E were normalized to those shown in panel B.