Figure 4.

Continuous RSC activity is necessary for formation of a stable transfer product. Dimer transfer reactions (A) and octamer transfer reactions (B) using the substrates from Figure 1 at 1 mM ATP were analyzed on a native gel. The left gels show the reaction with no ADP added and the right gels show reactions in which excess ADP was added after 3 min. The gels were scanned in the Cy5 channel. The graph shows the quantification of transfer for the reactions with no ADP added as black circles with a solid black line and ADP added at 3 min as open triangles with a dashed line. In (A) the values for dimer transfer are calculated from three independent experiments; error bars show the standard error of the mean. For octamer transfer one representative experiment is shown. Other experiments where ADP was added at different time points showed the same requirement for ATP (data not shown). In (B), left panel, all the time-points shown are from the same gel but lanes between the 3 min and 20 min time-points were omitted to enable a direct visual comparison with the gel in the right panel