Extended Data Fig. 9: Fig. S8. Target cleavage by EfPiwi and Siwi (part 2).
a, Guide-target pairing schematic for targets with varying degrees of complementarity to piRNA 3' end used in Fig. 4a and b, e. b, Quantification target RNAs (1 nM) cleaved after treating with excess (100 nM) EfPiwi loaded with a 22 or 25 nt guide or hAGO2 loaded with a 22 nt guide for 1 h. n = 3 independent experiments, data are mean ± s.d. c, Guide-target pairing schematic for targets with 3 nt mismatched regions. d, Quantification mismatched target RNAs (1 nM) cleaved after treating with excess (100 nM) EfPiwi loaded with a 22 or 25 nt guide or hAGO2 loaded with a 22 nt guide for 1 h. n = 3 independent experiments, data are mean ± s.d. e, Cleavage of targets with varying degrees of complementarity to piRNA 3' end (shown panel a) by Siwi (2 mM MnCl2, 37 °C). Gel is representative of n = 3 independent experiments, with data plotted as mean ± s.d. shown below. f, From Fig. 4c: sequences of the 26 target RNAs, with 0–8 mismatches opposite piRNA nucleotides g11–g18, that were most readily cleaved by EfPiwi (listed in order of cleavage product abundance). Mismatched nucleotides coloured yellow. Greyed out sequences indicate constant regions shared by all target RNAs. Triangle indicates cleavage site.
