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. 2021 Mar 5;12:1484. doi: 10.1038/s41467-021-21611-8

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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. 5 — a Confocal micrographs of GFP expressing dopaminergic neurons with and without injury and vhp-1 RNAi. Shown are average or maximum projections of Z-stacks. Reference Fig. 5b, c. Scale = 50 µm. b Quantification of CEP neuronal phenotype from vhp-1 RNAi maximum projections. vhp-1 RNAi uninjured (Un) versus injured *p = 0.0124, ***p = 0.0003. vhp-1 RNAi versus empty vector (EV, dashed lines) 1 and 48 hour (h) post injury, ns (not significant) p = 0.8122, and ****p < 0.0001 respectively, chi-squared test. c Large-particle flow cytometry time-course comparing effects of dopaminergic GFP retention in injured vhp-1 RNAi (green squares) to uninjured vhp-1 RNAi (---). Injured empty vector (EV, blue squares) reference from Fig. 3D, gray dashed lines. Mean ± 95% CI. Mixed-effects analysis with Tukey’s multiple comparisons test. See Supplementary Data 5 for statistics. d Nose-touch response of uninjured (tan upright triangles) or injured (green inverted triangles) worms on vhp-1 RNAi. Injured empty vector (EV, blue squares) reference from Fig. 1J (gray dashed lines) Mean ± SEM. See Supplementary Data 5 for statistics. e Chemotaxis of uninjured (Un, black and tan) or injured (Inj, blue and green) worms on empty vector (EV, black and blue) or vhp-1 RNAi (tan and green) with butanol 48 h post injury. Chemotactic index of 1.0 indicates 100% preference for butanol and 0.0 indicates no preference. Mean ± SEM, biological replicates denoted. See Supplementary Data 5 for statistics. f Large-particle flow cytometry results of wild type, PMD13, and vhp-1(sa366) mutants, PMD153, in green two days post trauma. Dopaminergic GFP index indicates relative loss of dat-1p::GFP signal in injured vhp-1(sa366) mutants (green) compared to injured wild type. Mean ± 95% CI. **p < 0.0015, two-sided paired t-test, n = 1902 for wild type and n = 1342 for vhp-1(sa366) over three independent trials. Positive values indicate reduced loss of fluorescence when compared to age-matched uninjured controls. A value of one indicates no change of fluorescence compared to uninjured controls.