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. Author manuscript; available in PMC: 2022 Dec 2.
Published in final edited form as: Mol Cell. 2021 Sep 29:S1097-2765(21)00746-2. doi: 10.1016/j.molcel.2021.09.012

Figure 2. Complementary RNAs Destabilize MIWI-loaded, Unmethylated piRNAs In Testis Lysate.

Figure 2.

(A) MIWI piRISC, loaded with a synthetic, 5′-32P-radiolabeled, 30-nt piRNA bearing a 2′-O-methyl or hydroxyl 3′ end was incubated in Henmt1em1/em1 testis lysate with 30-nt, fully 2′-O-methylated complementary RNA (cRNA). The mean of the guide abundance is shown (n = 3).

(B) As in (A), a time series is shown for MIWI piRISC incubated in Henmt1em1/em1 testis lysate with 30-nt, fully 2′-O-methylated cRNA.

(C) As in (A), except Henmt1em1/em1 testis lysate was preincubated for 5 min at the indicated temperature before adding MIWI piRISC and cRNA; MIWI piRISC was then incubated in lysate with cRNA for an additional 5 h.