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. 2023 Jul 24;14:4452. doi: 10.1038/s41467-023-40142-y

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a–h NMJs of MN4-Ib in third instar larvae of non-ablated and Is ablated larvae in control, glia shark knockdown, muscle shark knockdown, and double knockdown backgrounds, labeled with GFP (green) and HRP (magenta). NMJ expansion was observed upon Is MN ablation (b), which is blocked by glia shark knockdown (d) or double knockdown (h). i Cartoon representation of a dissected larva (pink) and a hemisegment highlighted by dashed red rectangle. The target muscle examined in this figure is shown in blue. Cartoon is generated with Biorender. j Quantification of MN4-Ib bouton number in non-ablated and Is ablated larvae in control, glial shark knockdown, muscle shark knockdown, and double knockdown backgrounds. Control (N = 22 and 23 NMJs), t(43) = 3.598, p = 0.0008, unpaired t test, two-tailed. Glial shark knockdown (N = 21 and 22 NMJs), t(41) = 1.566, p = 0.1250, unpaired t test, two-tailed. Muscle shark knockdown (N = 23 and 19 NMJs), t(40) = 3.220, p = 0.0025, unpaired t test, two-tailed. Double shark knockdown (N = 16 and 22 NMJs), t(36) = 0.3390, p = 0.7366, unpaired t test, two-tailed. k Quantification of normalized EPSP of Is ablated larvae in control, glia shark knockdown, muscle shark knockdown, and double knockdown backgrounds. F(3, 71) = 5.533, p = 0.0018, One-way ANOVA. Is ablated control vs glia shark knockdown, p = 0.0062. Is ablated control vs muscle shark knockdown, p = 0.0105. Is ablated control vs double knockdown, p = 0.0093. Is ablated control vs Ib/Ib+Is, t(28.04) = 4.485, p = 0.0001, unpaired t test, two-tailed, with Welch’s correction. Is ablated in glia shark knockdown vs Ib/Ib+Is, t(30) = 2.798, p = 0.0089, unpaired t test, two-tailed. Is ablated in muscle shark knockdown vs Ib/Ib+Is, t(30) = 2.329, p = 0.0268, unpaired t test, two-tailed. Is ablated in double knockdown vs Ib/Ib+Is, t(25) = 2.254, p = 0.0332, unpaired t test, two-tailed. l Quantification of normalized quantal content of Is ablated larvae in control, glia shark knockdown, muscle shark knockdown, and double knockdown backgrounds. F(3, 71) = 4.437, p = 0.0065, One-way ANOVA. Is ablated control vs glia shark knockdown, p = 0.0466. Is ablated control vs muscle shark knockdown, p = 0.9470. Is ablated control vs double knockdown, p = 0.0214. For (k, l), N (NMJs) = 20, 20, 20, 15. Error bars indicate ± SEM, ns = non-significant, *p < 0.05, **p < 0.01, ***p < 0.001.