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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 knockdown inhibits leucine sensing but has no impact on general IPC general function. L3 larvae were reared on standard medium until reaching the feeding L3 stage. Intracellular IPC Dilp2 stores were measured by Dilp2 immunolabeling in animals that were either starved or fed prior to brain dissection and Dilp2 labeling. Dilp2 is stored in IPCs of the control genotype (+>NaChBac) when the animals are starved. The low immunofluorescence level in fed animals reflects Dilp2 secretion upon feeding (right panels). Larvae that coexpress UAS-NaChBac and the JhI-21 knockdown constructs in the IPCs have low stores of Dilp2 even if they have been starved prior to the experiment (left panels). Statistics: ****p < 0.0001, ns > 0.05, Mann-Whitney test. Data are mean ± SEM.