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. 2018 Jan 30;8:1908. doi: 10.1038/s41598-018-20394-1

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© The Author(s) 2018

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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JhI-21 participates in autonomous leucine-sensing in IPCs. L3 larvae were reared on standard medium until reaching the feeding L3 stage and starved for 24 h. The brains of two control genotypes (Dilp2>+ and +>JhI-21^dsRNA) and one JhI-21 knockdown genotype (Dilp2 > JhI-21^dsRNA) were dissected and cultured overnight before fixation and Dilp2 immunolabeling. (a) Dilp2 immunolabeling was high in brains taken from starved larvae when cultured in regular Drosophila Schneider’s medium not supplied in leucine. (b) Dilp2 immunolabeling of control genotypes (grey bars) were low when brains were cultured in Drosophila Schneider’s medium that contained additional 20 mM of leucine. In IPCs in which JhI-21 expression was knocked down (Dilp2 > JhI-21^dsRNA, blue bar), internal Dilp2 stores remained high. Statistics in a: ns > 0.05, 1-Way-ANOVA followed by Bonferroni’s post hoc test, Statistics in b: ***p < 0.001, Kruskall-Wallis test followed by Dunn’s post hoc test. Data are mean ± SEM.