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. 2021 Feb 19;12:1160. doi: 10.1038/s41467-021-21429-4

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© The Author(s) 2021

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. 4 — a Brain midline labeled with anti-PDF at 5 DPE with cortex glia-driven draper-RNAi (CG, R54H02-Gal4 > draper-RNAi, left) or ensheathing glia-driven draper-RNAi (EG, R56F03-Gal4 > draper-RNAi, right). Boxed regions are the distal medial bundle (MDBL) or proximal subesophageal zone (SEZ). Scale bar: 25 μm. Images are representative of two independent experiments. b Quantification of anti-PDF area (top) or puncta (bottom) comparing MDBL and SEZ with CG > draper-RNAi (left, black/gray) and EG > draper-RNAi (right, blue/light blue). Area: CG > draper-RNAi, Two-sided Mann–Whitney, p = 0.0006, 26.61 ± 17.89 n = 7 MDBL, 94.62 ± 13.93 n = 8 SEZ. EG > draper-RNAi, Two-sided t test, p = 0.1084, 33.33 ± 6.844 n = 8 MDBL, 59.89 ± 13.89 n = 8 SEZ. Puncta: CG > draper-RNAi, two-sided Mann–Whitney, p = 0.0079, 4.28 ± 2.643 n = 7 MDBL, 18.88 ± 3.573 n = 8 SEZ. EG > draper-RNAi, Two-sided t test, p = 0.6049, 12.00 ± 2.360 n = 8 MDBL, 13.88 ± 2.642 n = 8 SEZ. c Brain slices from 5 DPE control (CG, R54H02-Gal4/+, top) and draper-RNAi (CG, R54H02-Gal4 > draper-RNAi) labeled with anti-PDF (green, left), nuclear anti-Draq5 (magenta, middle) and merge (left). Arrows indicate colocalized ant-PDF and -Draq5 (PDF-Tri soma). Arrowhead indicates PDF-Tri neuron slightly out of plane. Right: Quantification of PDF-Tri neuron cell bodies per brain. Two-sided Mann–Whitney, p = 0.0009, 0.2308 ± 0.1662 n = 13 control, 1.643 ± 0.3249 n = 14 draper-RNAi. d Brain slices from 5 DPE control (EG, R56F03-Gal4/+, top) and draper-RNAi (EG, R56F03-Gal4 > draper-RNAi) labeled with anti-PDF (green, left), nuclear anti-Draq5 (magenta, middle) and merge (left). Scale bar: 25 μm. Right: Quantification of PDF-Tri neuron cell bodies per brain. Two-sided Mann–Whitney, p = 0.1567, 0.000 ± 0.000 n = 16 control, 0.1818 ± 0.1220 n = 11 draper-RNAi. Scatter plots show mean ± SEM. Box and whisker plots show quartiles with max and min values, with plus sign indicating the mean. Sample size is n = number of animals. Significance: p > 0.05 (not significant N.S.), p < 0.01 (**), and p < 0.001 (***). Source data for this figure are provided in Source Data file.