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. 2016 Oct 20;6:35546. doi: 10.1038/srep35546

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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To identify key genes involved in regulation of Vg gene expression, dsRNA targeting key genes in ecdysone (Phantom, Shade, EcR, USP, BR-C), JH (Met, SRC, Kr-h1) and nutrition (mTOR, cTOR, GATAa, InR1, InR2, ILP1 and ILP2) signaling pathways were injected into the virgin adult females. The insects were incubated for five days after which they were blood fed and mated. Two days after feeding and mating, the total RNA was isolated and used to determine relative levels of Vg1, Vg2 and Vg3 mRNA levels using RPL8 gene expression for normalization. Each bar represents the average and standard deviation of three biological replicates. Student t-test was performed (p ≤ 0.05). All experiment were repeated twice.