Fig. (4) NS5A does not activate the NS5B replication reaction with the 3’ END (-) template. A. Radiolabeled 3’ END (-) template. B. NS5B (2.5 µM) replication reaction with the X(+) template (lane 1) or the 3’ END (-) template (2.5 µM each) C. NS5B (0.3 µM, lane 1, 2) plus or minus pre-incubated NS5A at 10 pM (lane 2). NS5B (0.2 µM, lane 3, 4) plus or minus pre-incubated NS5A at 10 pM (lane 4). NS5B (0.1 µM, lane 5, 6) plus or minus pre-incubated NS5A at 10 pM (lane 6). The reaction proceeded for 2 hours at 30°C after addition of the template (0.2-0.5 µM), and NTPs. Arrow indicates the expected RNA product.