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. 2020 Feb 26;10:3464. doi: 10.1038/s41598-020-60340-8

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Effects of RNAi knockdown of MCO genes on nymphs of P. stali. (A) Effects of RNAi on third instar nymphs, in which dsRNA for each MCO gene was injected into newly molted third instar nymphs. Note that knockdown of PsMCO2 led to high mortality during molting. (B) Phenotypes of newly molted fourth instar nymphs that were injected with either EGFP dsRNA as a control (left panel) or PsMCO2 dsRNA (right panel). All the PsMCO2 dsRNA-injected insects were not tanned. (C) Effects of RNAi on fifth instar nymphs. All the PsMCO2 dsRNA-injected insects died during molting. (D) A fifth instar nymph (left), a 1-day-old adult that were injected with EGFP dsRNA as a control (middle), and a typical image of molting failure induced by injection with PsMCO2 dsRNA (right). PsMCO2 RNAi causes incomplete molting. Arrowheads indicate nymphal (red) and adult (green) cuticles. Numbers above bars show sample sizes (i.e. biological replicates) in. (A,C) Scale bars show 5 mm in (B,D).