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. 2017 Nov 27;7:16395. doi: 10.1038/s41598-017-16625-6

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© The Author(s) 2017

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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RT-qPCR analysis of the expression profiles of the proteins relates to cuticle formation. P_< 0.5: <0.5 hr after injection; P₁₂–P₄₈: 12–48 hrs after pupation. The relative expression levels are expressed as means ± SD (n = 3), with uninjected pupae at <0.5 hr after injection as the calibrator. Asterisks indicate significant differences (*p < 0.005; **p < 0.001) from the relative mRNA levels of uninjected control at each time-point. (a) The expression profiles of eight proteins related to cuticle formation in uninjected control, GFP dsRNA-injected control and Aacec B dsRNA-injected pupae. AaPPO 1: Ae. aegypti prophenoloxidase 1; AaPPO 3: Ae. aegypti prophenoloxidase 3; AaPPO 4: Ae. aegypti prophenoloxidase 4; CBP: chitin binding protein; CP: cuticle protein; 78E: pupal cuticle protein 78E; SP: serine protease; Serpin: serine protease inhibitor. (b) RT-qPCR analysis of AaPPO 1, 3 and 4 expression in uninjected control, GFP dsRNA-injected control and AaPPO 3 dsRNA-injected pupae.