Fig. (5) NS5A activation of NS5B occurs during elongation on a synthetic template. A. Replication reaction with the LTMOA template (1 µM) in the absence (lanes 1 and 2) or presence (lanes 3 and 4) of NS5A (0.2 pM) for 1 (lanes 1 and 3) and 2 hours (lanes 2 and 4) B. NS5B-containing complexes on LTMOA can be stalled at G21 (lane 1), then released into elongation (lanes 2-5) for 20, 70, 130 and 310 seconds. To quantify the formation of RNA intermediates, the gel was divided into seven regions (R1-R7) for each time point. C. Quantitation of an experiment as in 5B with timepoints of 10 (panel 2), 20 (panel 3), 30 (panel 4), and 90 (panel 5) seconds with and without NS5A.