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. 2018 Feb 28;9:872. doi: 10.1038/s41467-018-02873-1

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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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Fig. 3 — Virtually normal locomotor and odor avoidance behaviors for 30 days after surgery. a Behavioral chamber for locomotor and odor avoidance assays, with odor delivery inlets at both sides. Red box: image region-of-interest analyzed to track fly movements. Scale bar: 10 mm. b Fly tracking steps: (1) Isolate the region-of-interest; (2) Subtract the mean background image; (3) Group pixels into background (black) and foreground/object (white) pixels using Otsu’s method and filter objects by size to isolate the fly; (4) Compute fly’s current centroid. c An example of walking trajectory (left) and speed trace (right). Repulsive odor, 4-methyl-cyclohexanol (3% in air), entered first from the left and then the right inlet. Pink indicates these intervals and entry routes; the opposite inlet delivered fresh air (gray shading). Otherwise, both inlets delivered air (white). d Flies that had the full surgery or glue on the cuticle had indistinguishable activity indices from control flies on Days 6 (P = 0.9) and 10 (P = 0.6) (Kruskal–Wallis ANOVA; 10 flies per group). On Days 2 (P = 0.03) and 20 (P = 0.04), the groups exhibited significant differences (n = 8–10 flies per group; Kruskal–Wallis ANOVA), which were insufficient to yield significant paired post-hoc tests between groups (P = 0.06–0.9; U-test with Holm–Bonferroni correction). On Day 30, Kruskal–Wallis ANOVA revealed significant differences between groups (P = 0.02, n = 7–10 flies); post-hoc testing showed surgery group flies had lower activity indices than control flies (P = 0.02; U-test with Holm–Bonferroni correction). *P < 0.05. e Flies that had surgery or glue on the cuticle had indistinguishable mean locomotor speeds from control flies on Days 6–20 (P = 0.1–0.9; Kruskal–Wallis ANOVA; n = 8–10 flies per group). On Days 2 and 30, the groups exhibited significant differences (P = 0.03, Day 2, P = 0.04, Day 30; Kruskal–Wallis ANOVA; n = 7–10 flies per group), which were insufficient to yield significant paired, post-hoc tests between groups (P = 0.05–0.6; U-test with Holm–Bonferroni correction). f, g Maximal walking speeds (f) and odor avoidance indices (g) were indistinguishable between groups for 30 days (P = 0.2–0.9 for all comparisons of walking speeds and odor avoidance indices; n = 7–10 flies per group; Kruskal–Wallis ANOVA). Error bars: s.e.m.