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. 2022 Jul 2;13:3814. doi: 10.1038/s41467-022-31253-z

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© The Author(s) 2022

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 Immunostaining of TMEM63::GFP (anti-GFP) labels the sensory cilia of Or42b neurons. White dashed line represents the border of a single Or42b neuron, with the main anatomical features shown. Each result was repeated three times. Scale bar, 10 μm. b Spatial distribution of control (upper left), Tmem63^KO (upper right), DmTMEM63-Rescue (lower left) and HsTMEM63B-Rescue (lower right) flies assayed in a 20% to 70% RH gradient. The yellow dashed circle denotes the area above 20% RH, and the green circle indicates the 70% RH region. n = 25 flies for each group. c The time course indicating the attraction indexes of heterozygous and homozygous Tmem63^KO flies assayed in the arena with a 20% to 70% RH setting. Tmem63^KO/+, n = 8 assays; Tmem63^KO, n = 12. d The time course indicating the attraction indexes of UAS Ctrl, DmTMEM63-Rescue and HsTMEM63B-Rescue flies assayed in the arena with a 20% to 70% RH setting. n = 9, 11, 11 assays. Genotypes are UAS Ctrl: Tmem63^KO; UAS-DmTMEM63/+, DmTMEM63-Rescue: Tmem63^KO; UAS-DmTMEM63/Or42b-Gal4 and HsTMEM63B-Rescue: Tmem63^KO; UAS-HsTMEM63B/Or42b-Gal4. e Probability distributions of distance between flies and moist center during 0–10 s (blue) and 80–90 s (red) after the onset of the assay. n = 25, 25, 24, 25 flies. Each result was reproducible in three independent experiments. f Attraction indexes of flies with indicated genotype tested in the 20–70% RH condition. n = 14, 12, 9, 8, 10, 7, 7, 6 assays. One-way ANOVA followed by Dunnett’s test for multiple comparison with the control group. g Expression of Drosophila TMEM63 or human TMEM63B in Or42b-Gal4 neurons with the Tmem63^KO allele restored the attraction behavior to 70% RH. n = 10, 9, 11, 12, 11 assays. Two-tailed unpaired t test. For (c, d), data points are mean values and shaded area represents ±SEM. For (f, g), data are mean ± SEM. Source data are provided as a Source Data file.