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. 2024 Jan 10;15:447. doi: 10.1038/s41467-024-44732-2

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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 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. 1 — a Micrography GFAP staining of CTR and DJ1 KO day 40, 100, and 200 midbrain organoids. b Midbrain differentiation and astrocytes extraction protocol schematic. c Immunoblots for α-syn, phospho-α-syn (S129) (n = 3, Two-tailed t-test), and corresponding loading controls actin (ACTB) and GAPDH for CTR and DJ1 KO day 40 (α-syn n = 3, phospho-α-syn (S129), n = 3) and day 200 midbrain organoids (α-syn n = 3; phospho-α-syn (S129), n = 6). d Dot blots for oligomeric α-syn and actin (ACTB)/Ponceau loading control for CTR and DJ1 KO day 40 TTX soluble (n = 6); SDS soluble fractions (n = 6) and day 200 TTX soluble (n = 6) and SDS soluble fractions (n = 3) in midbrain organoids. e Immunoblots for LC3 I/II, P62, actin (ACTB) loading control in BAF – and + treated CTR and DJ1 KO day 100 midbrain organoids, graphs report LC3 I/II basal (n = 6), LC3 I/II flux (n = 6), P62 basal (n = 6), P62 flux (n = 6). f Immunoblots for GBA and ACTB/GAPDH loading control for CTR and DJ1 KO day 40 (n = 6) and day 200 (n = 3) midbrain organoids. All data are represented in mean ± S.E.M, data points are individual well differentiation, and the p-value was reported on the graph highlighted comparison. For all the comparisons, a Two-tailed t-test was applied. Panels c and f share the same loading controls. All measurements were taken from distinct samples.