Figure 3.

Single-molecule imaging of the target cleavage reaction. (A) Schematic representation of the single-molecule imaging. 3′-Cy3-labeled target RNA (green target) was tethered on the glass surface and incubated with S2 cell lysate containing Ago2-RISC with 3′-Cy5-labeled guide strand (red RISC) in the presence or absence of CrPV-1A. (B) Time-lapse single-molecule imaging of target cleavage with or without CrPV-1A. Yellow circles denote co-localization of the green target and the red RISC spots. (C) Quantification of the time-lapse single-molecule experiment. The number of green spots decreased remarkably in the absence of CrPV-1A (– CrPV-1A), but not in the presence of CrPV-1A (+ CrPV-1A) or without RISC assembly (– siRNA). Error bars indicate the SD from three independent experiments.