Extended Data Fig. 8: Target cleavage by EfPiwi and Siwi (part 1).
a, Denaturing gels showing cleavage of g2-g21 matched 32P-labeled target RNA by EfPiwi, hAGO2 or Siwi in the presence of various divalent cations (2 mM each). Schematic of piRNA-target pairing shown (top). Gels are representative results for n = 3 independent experiments for EfPiwi and hAGO2, and n = 2 independent experiments for Siwi. b, Time course showing cleavage of g2-g21 paired 32P-labeled target RNA by EfPiwi in the presence of Mg2+, Mn2+, or both at approximate physiological divalent cation concentrations. c, Cleavage of g2-g21 matched 32P-labeled target RNA by EfPiwi at various temperatures shows activity over the full physiological range (17–30 °C). Gel is representative of n = 3 independent experiments. d, Quantification of results (and replicates) in c. e, Cleavage of target RNAs with varying degrees of 3' complementarity by EfPiwi at 30 °C. In b, d and e, n = 3 independent experiments, data are mean ± s.d.
