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. 2021 Sep 22;11:18807. doi: 10.1038/s41598-021-98334-9

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Tc-Vn and Tc-Spi activate redundantly the larval-pupal transition in Tribolium. (A) Tc-vn mRNA levels measured by qRT-PCR in penultimate (L6) and ultimate (L7) instar larvae. Transcript abundance values are normalized against the Tc-Rpl32 transcript. Fold changes are relative to the expression of Tc-vn in day 0 of L6 larvae, arbitrarily set to 1. Error bars indicate the SEM (n = 5). (B) L6 larvae were injected with dsMock (Control), dsTc-spi (Tc-spi^RNAi) or both dsTc-spi and dsTc-vn simultaneously (Tc-spi^RNAi + Tc-vn^RNAi) and left until the ensuing molts. Ventral views of Control and Tc-spi^RNAi pupae and a Tc-spi^RNAi + Tc-vn^RNAi larva arrested at the larval-pupal transition. Scale bar represents 0.5 mm. (C) Transcript levels of Tc-Hr3, Tc-E75 and TcBr-C, (D) Tc-E93 and (E) biosynthetic ecdysone genes Tc-phm, Tc-dib, Tc-spok and Tc-shd measured by qRT-PCR in 5-day-old Control, and Tc-spi^RNAi + Tcvn^RNAi L7 larvae. Transcript abundance values were normalized against the Tc-Rpl32 transcript. Average values of three independent datasets are shown with standard errors (n = 7). Asterisks indicate differences statistically significant at *p < 0.05 and **p < 0.001 (t-test) ***p < 0.0001 (t-test).