Fig. 4 Dcr-2 binds to the 3′UTR of Toll mRNA. Dcr-2 binds to the 3′UTR of Toll mRNA. (A) Cultured S2 cells were transfected with empty vector (lanes 1 and 2), the plasmid expressing Dcr-2–Flag (lanes 3 and 4), and in vitro–transcribed Toll 3′UTR (lanes 2 and 4), as indicated. Crude cell lysates were prepared and subjected to RNA-IP using anti-Flag antibody. RNA extracts prepared from precipitates and total cell lysates were subjected to Northern blots, as indicated. Precipitates and total cell lysates were also subjected to Western blots using the indicated antibodies. (B) RNA extracts were prepared from RNA-IP precipitates as in (A) and subjected to qRT-PCR of Toll mRNA (n = 3; *P < 0.01, two-tailed Student’s t test; error bars, SD). (C) RNA extracts were prepared as described in (B) and subjected to RT-PCR with Toll-specific primers. RT-PCR products were analyzed via agarose gel electrophoresis. NB, Northern blots; IB, immunoblots or Western blots. Zhaowei Wang et al. Sci Adv 2015;1:e1500228 Copyright © 2015, The Authors