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. 2022 Jun 7;13:3271. doi: 10.1038/s41467-022-30869-5

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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 — Quantification (a) of individual fmo-2p::mCherry worms on fed (pink), and DR (blue) treated with 100 µM mianserin (purple), 100 µM thioridazine (orange), or combined (yellow) worms fed vector or ser-4 RNAi. n = 18 (vector fed), 23 (vector DR), 25 (vector fed + mianserin), 15 (vector fed + thioridazine), 19 (vector fed + mianserin + thioridazine), 16 (ser-4 RNAi fed), 16 (ser-4 RNAi DR), 11 (ser-4 RNAi fed + mianserin), 13 (ser-4 RNAi fed + thioridazine), 10 (ser-4 RNAi fed + mianserin + thioridazine) biologically independent animals. p-value = 3.47e-07 (vector fed vs. vector DR), 8.12e-19 (vector fed vs. vector fed + mianserin), 5.91e-08 (vector fed vs. vector fed + thioridazine), 1.30e-06 (vector fed vs. vector fed + mianserin + thioridazine), 5.16e-10 (ser-4 RNAi fed vs. ser-4 RNAi DR), 0.04998 (ser-4 RNAi fed vs. ser-4 RNAi fed + mianserin), 0.126 (ser-4 RNAi fed vs. ser-4 RNAi fed + thioridazine), 0.0165 (ser-4 RNAi fed vs. ser-4 RNAi fed + mianserin + thioridazine). Survival curves (b) of WT animals on vector RNAi in pink and ser-4 RNAi in purple on fed (solid lines) or DR (dotted lines) conditions. Images (c) and quantification (d) of fmo-2p::mCherry or ser-4 + /- tissue-specific rescue on fed (pink) and DR (blue). Scale bar, 1 mm. n = 10 (control fed), 14 (control DR), 14 (ser-4(-) fed), 8 (ser-4(-) DR), 10 (ser-4(-) + intestinal fed), 12 (ser-4(-) + intestinal DR), 9 (ser-4(-) + neuronal fed), 17 (ser-4(-) + neuronal DR) biologically independent animals. p-value = 3.10e-08 (control DR vs. ser-4 (-) DR), 5.19e-10 (control DR vs. ser-4 (-) + intestinal DR), 0.131 (control DR vs. ser-4 (-) + neuronal DR). Images (e) and quantification (f) of fmo-2p::mCherry or ser-4 KO + /- GABAergic rescue on fed (pink), DR (blue) and DR + food smell (purple). Scale bar, 1 mm. n = 14 (WT fed), 21 (WT DR), 18 (WT DR + food smell), 22 (ser-4(-) fed), 17 (ser-4(-) DR), 21 (ser-4(-) DR + food smell), 11 (ser-4(-) + GABA rescue fed), 31 (ser-4(-) + GABA rescue DR), 23 (ser-4(-) + GABA rescue DR + food smell) biologically independent animals. p-value = 1.72e-05 (WT DR vs. WT DR + food smell), 0.849 (ser-4(-) DR vs. ser-4(-) DR + food smell), 4.00e-08 (ser-4(-) + GABA rescue DR vs. ser-4(-) + GABA rescue DR + food smell). Quantification (g) of individual fmo-2p::mCherry worms on fed (pink), and DR (blue) treated with 100 µM mianserin (purple), 100 µM thioridazine (orange), or combined (yellow) worms fed vector or dop-3 RNAi. n = 23 (vector fed), 29 (vector DR), 6 (vector fed + mianserin), 14 (vector fed + thioridazine), 34 (vector fed + mianserin + thioridazine), 24 (dop-3 RNAi fed), 23 (dop-3 RNAi DR), 26 (dop-3 RNAi fed + mianserin), 15 (dop-3 RNAi fed + thioridazine), 21 (dop-3 RNAi fed + mianserin + thioridazine) biologically independent animals. p-value = 1.69e-16 (vector fed vs. vector DR), 0.0268 (vector fed vs. vector fed + mianserin), 8.34e-07 (vector fed vs. vector fed + thioridazine), 1.03e-07 (vector fed vs. vector fed + mianserin + thioridazine), 1.88e-08 (dop-3 RNAi fed vs. dop-3 RNAi DR), 0.542 (dop-3 RNAi fed vs. dop-3 RNAi fed + mianserin), 0.166 (dop-3 RNAi fed vs. dop-3 RNAi fed + thioridazine), 0.00648 (dop-3 RNAi fed vs. dop-3 RNAi fed + mianserin + thioridazine). Survival curves (h) of WT animals on vector RNAi in pink and dop-3 RNAi in purple on fed (solid lines) or DR (dotted lines) conditions. ****P < .0001 (Welch Two Sample t-test, two-sided). ^###P < 0.001 when compared to ser-4 (-) or ser-4 (-) + intestinal fed (Welch Two Sample t-test, two-sided); ^#P < 0.05, ^####P < 0.0001 when compared to ser-4/dop-3 RNAi fed (Welch Two Sample t-test, two-sided). The box plots display the median by the middle line of the box. The upper boundary of the box indicates the 75% interquartile range, and the lower boundary indicates the 25% interquartile range.