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. 2021 Jan 21;11:2061. doi: 10.1038/s41598-021-81548-2

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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 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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Knockdown of Dα1 or Dβ2 in the clock bearing cells disrupts circadian rhythmicity and sleep with no further effect by addition of neonicotinoids. Representative actograms for (a) Dα1 knock down (tim-Gal4 > uas-nAChR-Dα1) and (b) Dβ2 knockdown (tim-Gal4 > uas-nAChR-Dβ2. Effects of knocking down Dα1 in clock bearing cells on (c) rhythmicity (RS) (F_2,79 = 11.8, p < 0.001), (d) number (no.) of sleep episodes in day (F_2,79 = 2.9, p = 0.063) and night (F_2,79 = 12.3, p < 0.001) and (e) mean episode length in day (F_2,79 = 5.1, p = 0.008) and night (F_2,79 = 8.3, p = 0.001). Effects of knocking down Dβ2 in clock bearing cells on (f) rhythmicity (F_2,79 = 31.5, p < 0.001), (g) no. of sleep episodes in day (F_2,79 = 1.6, p = 0.211) and night (F_2,79 = 28.2, p < 0.001) and (h) mean episode length in day (F_2,79 = 11.2, p < 0.001) and night (F_2,79 = 9.4, p < 0.001). Each data point represents a single fly, n = 19–32 flies per treatment. There was no additive effect of 50 µg/L of IM and CLO on (i) tim > α1 (F_2,85 = 0.4, p = 0.677) and (j) tim > β2 (F_2,89 = 1.1, p = 0.336). Each data point represents a single fly, n = 24–32 flies per treatment.