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. 2020 Apr 9;10:6147. doi: 10.1038/s41598-020-63169-3

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© The Author(s) 2020

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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trans-Tango downstream targets of PD2a1/b1, MBON- 𝛾2α’1 and MBON- β’2mp neurons. (a) R37G11-GAL4-driven myrGFP (green) labels 6-7 cells in the LH, whereas the trans-Tango signal (magenta) appears to be strongest in overlapping neuropil regions and in random sparse KC subsets in the MB (boxed area enlarged in a’ and a”). (a’) R37G11-GAL4-driven myrGFP (green) labels 6-7 cells (arrowhead) in the LH (stippled). (a”) R37G11-GAL4-driven trans-Tango signal (magenta) marks cells (arrowhead) in the LH (stippled). (b) Confocal stack of MB (stippled), highlighting the PD2a1/b1 neurites in proximity to the vertical lobe and a trans-Tango signal in all sublobes, i.e., αβ, αβ’, and 𝛾. (c) Confocal stack of MB calyx (stippled) showing innervation by R37G11-GAL4-driven GFP (green) and trans-Tango signal in both KC somata and the calyx neuropil. (d) Confocal stack of left hemibrain highlighting R37G11-GAL4-driven myrGFP (green) and postsynaptic trans-Tango signal in layer 6 of the FSB (stippled). (e) MB077B-GAL4-driven myrGFP (green) labels 3-4 MBON-𝛾2α’1 neurons, whereas a fairly restricted putative downstream trans-Tango signal (magenta) most prominently labels MBON- β’2mp (boxed area enlarged in e’). (e’) Confocal stack of left hemibrain highlighting MB077B-GAL4-driven mCD8::GFP (green) and postsynaptic trans-Tango signal (magenta) in β’2mp of the MB and in putative LHON neurites (arrowheads). (f) MB011B-GAL4-driven myrGFP (green) labels MBON- 𝛾5β2α and MBON- β’2mp, whereas the putative downstream trans-Tango signal (magenta) labels several different cell types (boxed area enlarged in f’). (f’) Confocal stack of right hemisphere highlighting the postsynaptic signal of MBON- β’2mp in PD2 LHONs (arrowhead) in the LH (stippled) and in layer 4 & 5 of the FSB (stippled). (a–f’): Confocal stack views of 1-2 µm-thick optical section data. Scale bars: a,e,f: 50 µm; a’,a”,b-d,e’,f’: 20 µm.