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. 2009 Jul 22;284(38):26017–26028. doi: 10.1074/jbc.M109.010835

FIGURE 2.

FIGURE 2.

Cleavage activities for human Ago2 containing various length antisense RNAs. A, denaturing PAGE of 40-nucleotide sense RNA digested with GST-Ago2 containing the antisense RNAs shown in C, ranging in length from 17 to 25 nucleotides. B, denaturing PAGE of 40-nucleotide sense RNA digested with HA-Ago2 containing the antisense RNAs shown in C. Antisense RNA was incubated with Ago2 for 1 h prior to incubation with the 40-nucleotide sense RNA for an additional 1 h. T1, 40-nucleotide sense RNA digested with RNase T1 for 10 min at 24 °C. The positions of the T1 digestions adjacent to guanidine residues are shown next to the ladder. Arrows indicate the position of the human Ago2 cleavage site. C, sequences of the antisense RNAs with the corresponding lane designations. The 19-nucleotide antisense RNA contained either a 5′-phosphate (p) or 5′-hydroxyl (OH). The 19-nucleotide antisense RNA containing a 5′-phosphate corresponds to the antisense RNA in Fig. 1. D, percent sense RNA cleaved with GST-Ago2 (solid bar) or HA-Ago2 (hatched bar). The standard errors reported for the Ago2 cleavage activities are based on three experiments. The antisense RNA sequences tested and corresponding lane designations are described in C.