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. 2021 Feb 10;12:624165. doi: 10.3389/fphys.2021.624165

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Copyright © 2021 Leyria, El-Mawed, Orchard and Lange.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of dsRNA treatment on Rhopr-TRET mRNA expression. (A) Experimental scheme for (B–E). Panels (B,D) show the knockdown efficiency of dsRhopr-IR1 and dsRhopr-AKHR treatments, respectively. Panels (C,E) show the effects of down-regulation of Rhopr-IR1 and Rhopr-AKHR transcripts on Rhopr-TRET expression. (F) Experimental scheme for (G–J). Panels (G,I) show the knockdown efficiency of dsRhopr-IR1 and dsRhopr-AKHR treatments, respectively. Panels (H,J) show the effects of down-regulation of Rhopr-IR1 and Rhopr-AKHR transcripts on Rhopr-TRET mRNA expression following P_In (H), Rhopr-AKH (J) or saline stimulation. Transcript expression was tested by RT-qPCR. The y-axis represents the fold change in expression relative to dsARG injection after saline treatment (dsARG/S, value ∼ 1) obtained viageometric averaging using 18S ribosomal RNA subunit and β-actin as reference genes. The results are shown as the mean ± SEM (n = 4–5, where each n represents an individual tissue from 1 insect). Statistically significant differences were determined by Student’s t-test in all graphs except for (H,J) in which a one-way ANOVA and Tukey’s test as post hoc test was used (different letters indicate significant difference at P < 0.05). dsARG/S: dsARG insects with saline injection; dsARG/P_In: dsARG insects with porcine insulin injection: dsARG/Rhopr-AKH: dsARG insects with Rhopr-AKH injection; dsRhopr-IR1/S: dsRhopr/IR1 insects with saline injection; dsRhopr-IR1/P_In: dsRhopr-IR1 insects with porcine insulin injection; dsRhopr-AKHR/S: dsRhopr-AKHR insects with saline injection; dsRhopr-AKHR/Rhopr-AKH: dsRhopr-AKHR insects with Rhopr-AKH injection.