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. 2014 Apr 9;289(23):16270–16277. doi: 10.1074/jbc.M114.569707

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Effects of the order-of-addition on inhibition of RNase H activity. A, sequence of the polymerase-dependent substrate used in this assay. Primary RNase H cleavages are indicated. * indicates 3′-radiolabeled primer. B, graphic representation of the effects of the order-of-addition of reaction components in the absence of inhibitor. Percent activity refers to the percentage of RNA template converted into the −18 and −19 major products. C, effects of the order-of-addition of reaction components during inhibition by GSK5750 (0.5 μm) under pre-steady-state conditions. D, results of extended time course experiment using the inhibited pre-formed E-I-S complex from C.