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. 2024 Jan 4;15:205. doi: 10.1038/s41467-023-44233-8

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© The Author(s) 2024

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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Fig. 5 — Representative images (of i) of neurons immunostained for Vapa (white) and mitochondria (Mito-DsRed, red) (a); and Vapb (white) and mitochondria (Mito-DsRed, red) (d). Arrows depicting dendrites and arrowheads depicting axons, straightened and magnified for better visualization in (b–f). Representative line profiles (of i) showing local enrichment of Vapa near dendritic (b, arrow in a) and axonal mitochondria (c, arrowhead in a) and Vapb near dendritic (e, arrow in d) but not near axonal mitochondria (f, arrowhead in d). Map2 (cyan) was used to trace the dendrite, and the Vapa and Vapb signal was used to trace the axon (white lines). Vapa and Vapb antibodies were validated in Vapa and Vapb knockdown neurons, respectively. Scale bar, 5 μm. Representative correlation (of i) between individual fluorescent pixel intensities of mitochondria (Mito-DsRed) and Vapb show a strong correlation in the dendrite (g) but not in the axon (h). i The average correlation coefficient measured shows a strong correlation between mitochondria and Vapa (A), Vapb (B) in fixed dendrites, and Vapb in live dendrites (B_live, Vapb-emerald), compared to shuffled control (Shf, see Methods). In axons, a strong correlation between mitochondria and A was observed, but not between mitochondria and B and B_live compared to Shf. Furthermore, the correlation coefficient between Vapb (B and B_live) and mitochondria in dendrites is statistically significant compared to that in axons. n in dendrites, animals: 11, 2 (A), 7, 3 (B), 12, 4 (B_live), 14, 4 (Shf). n in axons, animals: 9, 2 (A), 8, 2 (B), 5, 1 (B_live), 11, 3 (Shf). One-way ANOVA, Tukey test, p-values: 0.00261 (B den versus ax), 0.01644 (B_live den versus ax), <0.0001 (A, B, B_live den versus Shf den), <0.0001 (A ax vs Shf ax), 0.00338 (A ax vs B and B_live ax). As a positive control, the correlation between two different fluorescent tags targeted to the mitochondrial matrix (Mito-DsRed and Mito-PAGFP) was used, which provided the upper limit of the correlation coefficient. As a negative control, the correlation between a mitochondrial (Mito-DsRed) and a non-mitochondrial tag was used for the lower limit of the correlation coefficient. Source Data files are provided.