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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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Exogenous Aacec B peptide is able to elevate the transcription levels of AaPPO 3, and Aacec B is able to be detected in the cytoplasm and nucleus of pupal cells. (a) The expression profile of AaPPO 3 in uninjected control, ddH₂O-injected control, 1 ng or 10 ng of Aacec B peptide was injected into pupae. 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. (b,c) Western blot analysis using antibodies specific for Aacec B: (b) cytoplasmic fraction and (c) nuclear fractions. β-actin and TATA-binding protein (TBP) were used as loading controls for the cytoplasmic and nuclear fractions, respectively. P_< 0.5: <0.5 hr after injection; P₁₂–P₄₈: 12–48 hrs after pupation. +: synthetic Aacec B peptide. Uncropped images are shown in Supplementary Figure S4.