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. 2019 Jan 10;10:124. doi: 10.1038/s41467-018-07929-w

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© The Author(s) 2019

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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Fig. 2 — RDL on CLPN^R82B09s mediates the inhibition from LRIN^R13B07s a–b. Optogenetic inhibition of LRIN^R13B07s allows excitation of CLPN^R82B09s. b is the statistics of peak calcium responses in a. The orange bar indicates the 540 nm light stimulation. The genotypes are R82B09-LexA/UAS-NpHR; LexAop-GCAMP6s/+ and R82B09-LexA/UAS-NpHR; R13B07-Gal4/ LexAop-GCAMP6s. c Expression pattern of R82B09-Gal4 in larval brain. Arrows point to cell bodies of CLPN ^R82B09s. d Morphology of single CLPN^R82B09 labeled by R82B09-Gal4. e Co-localization between the dendrites of CLPN^R82B09 and LRIN^R13B07 in larval brain. Yellow circle outlines the overlapping region between LRIN^R13B07 and CLPN^R82B09. f GRASP signal between CLPN^R82B09 and LRIN^R13B07. Yellow circle outlines verified contact region. g–i trans-Tango probes CLPN^R82B09 as immediate downstream of R13B07-Gal4 labeled neuron. g is the merged version of h and i. trans-Tango signal is in magenta. GFP signal driven by R82B09-Gal4 is in green. Arrows indicated the overlapping between trans-Tango signal and GFP signal that labels the cell body of a CLPN^R82B09. Inset in g shows the position of the co-localization in larval brain hemisphere. UAS-myrGFP,QUAS-mtdTomato(3xHA)/+;trans-Tango/+;R13B07-Gal4/R82B09-Gal4 is the genotype of the larva used. trans-Tango signals are reported by 3xHA. j–k Application of 100 mM GABA repressed the activation of CLPN^R82B09s by 100 μM acetycholine. h is the statistics of peak calcium responses in g. l–m Application of 100 μM RDL antagonist picrotoxin allows excitation of CLPN^R82B09s. j is the statistics of peak calcium responses in i. n–o Calcium response in CLPN^R82B09s before and after etomidate application. Arrow indicates application of drug. o is the statistics of peak responses in n. Scale bars, 20 μm in c–i. Samples in j–o were prepared from larvae of genotype R82B09-LexA;LexAop-GCAMP6s. Arrows indicate application of drugs in j, l, and n. *P < 0.05, **P < 0.01, ***P < 0.001, t-test in b, k, m, and o. Error bars, SEMs. Source data of a–b and j–o are provided as a source data file