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. 2018 Jun 8;9:2232. doi: 10.1038/s41467-018-04675-x

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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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Fig. 6 — Genetic ablation of larval LNs causes aberrant glomeruli organization. a Schematic of temperature shift. Larvae were transiently raised at 29 °C from late 3rd instar larvae to 6.5 h APF. b, d PN dendrites targeting to glomeruli DA1, VA1d, and DC3 of control and NP3056 > DTI (b), and control and 670 > DTI (d) brains. Adult brains were stained for Bruchpilot (magenta) to visualize individual glomeruli and GFP to visualize PN dendrites (green). c, e 3D reconstruction of ALs from control (top) and LN > DTI brains were shown in anterior view (left) and lateral view (right), respectively. Glomeruli DA1, VA1d, and DC3 are in green. f Bar graph showing the volume of UAS-DTI control, LN-GAL4 control, or LN > DTI ALs. g Ten variables (Methods) extracted from UAS-DTI control (gray dots), LN-GAL4 control (green (NP3056), magenta (449) or blue (670) dots outlined with black), and LN > DTI (green (NP3056), magenta (449), or blue (670) dots) ALs were subjected to principle component analysis (PCA). No obvious separation of control and LN > DTI ALs was observed. h The relative distance between any two glomeruli in NP3056-GAL4 control (left panel) and NP3056 > DTI (middle panel) ALs. The differences between a given pair of glomeruli in control and NP3056 > DTI ALs are shown in the right panel. i The differences between a given pair of glomeruli in control and 670 > DTI ALs. j (Left) The y-position of individual glomeruli in NP3056 control and NP3056 > DTI ALs. Glomeruli were aligned based on their mean y-position value in the GAL4 control ALs (Supplementary Note 4). Note that NP3056 > DTI glomeruli have larger y-position values. (Middle) The relative shift of individual glomeruli in NP3056 > DTI ALs was quantified as the deviation. Glomeruli were aligned as in the left panel. The mean deviation of all glomeruli is 14.0318 (black line). Gray lines indicate one standard deviation (SD) from the mean, which contains 68.3% of the glomeruli. (Right) A 3D-reconstructed AL shows glomeruli with deviation > 1 SD (pink) and <−1 SD (orange). k The y-position of individual glomeruli in 670 control and 670 > DTI ALs is shown. No obvious y-position shift was observed in 670 > DTI ALs. f, j, k Mean ± s.d