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. 2016 Nov 1;14(1):73–89. doi: 10.1080/15476286.2016.1251542

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© 2017 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 1. — RipSeq protocol retains BicD/Egl complex stability and enriches for known target mRNAs. (A-B) Extracts from Egl::GFP expressing embryos and wt control extracts were subjected to IP with anti-GFP antibodies. Western blotting tested for the presence of the BicD and Egl::GFP (A). Semiquantitative RT-PCR tested for the enrichment of known BicD/Egl mRNA targets and for the lack of enrichment of non-localizing mRNAs and house keeping mRNAs (B). (C) Rip-Seq enrichment of mRNAs from the test set. (D-E) Most top candidate BicD/Egl targets identified by Rip-Seq (D) could be validated by RT-qPCR analysis in an independent IP experiment (E). Error bars represent +/− SD of 2 independent IPs. Note that fold enrichment values of the 2 different experiments cannot be compared directly because they are calculated using different formulas (see methods). Nevertheless, enrichments in the Egl::GFP IPs relative to mock IPs is reproducible.