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. 2020 Jun 5;11:2835. doi: 10.1038/s41467-020-16696-6

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© The Author(s) 2020

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. 7 — a Loss of Rab35 protein expression in Schwann cell/DRG neuron co-culture explants from Rab35 WT and cKO^SC mouse embryos, One sample two-tailed Student’s t-test with a theoretical mean of 1, p < 0.0001, t = 25, df = 4, n = 3 WT, and n = 5 cKO^SC DRG pools of at least 10 explants each. b Representative confocal images of myelin-forming Schwann cell/DRG neuron co-culture explants from Rab35 WT, wild-type (Rab35^flox/flox) and cKO^SC mouse embryos either left untreated (NT) or treated with Rapamycin (RAPA). DAPI staining shows Schwann cell nuclei; NF-M staining, neurofilament, and Mbp, myelin basic protein to detect myelin segments. c Levels of p-S6 in WT and cKO^SC DRG explants following 2 and 5 days of ascorbic acid treatment in differentiating conditions. n = 1 independent experiment. d Quantification of the number of Mbp-positive myelin segments in co-culture explants from Rab35 WT and cKO^SC embryos either left untreated (NT) or treated with Rapamycin (RAPA): WT (NT): 27.18 ± 3.22, n = 8 DRG/coverslips; WT (RAPA): 54.51 ± 7.641, n = 9 DRG/coverslips; cKO^SC (NT): 15.38 ± 1.2, n = 11 DRG/coverslips; cKO^SC (RAPA): 32.79 ± 4.26, n = 9 DRG/coverslips, one-way non-parametric ANOVA followed by Dunn’s Multiple Comparison Test, p < 0.0001. e Representative confocal images and f quantification of the fraction of Mbp-positive fibers carrying myelin abnormalities in Rab35 cKO^SC explants either left untreated (NT) or treated with Rapamycin (RAPA). Asterisks mark Mbp-positive fibers carrying abnormalities. cKO^SC (NT): 32.62 ± 3.42, n = 330 fibers in n = 11 coverslips/DRG explants; cKO^SC (RAPA): 12.73 ± 1.89, n = 311 fibers in n = 9 coverslips/DRG explants, ***p = 0.0004, two-tailed nonparametric Mann–Whitney test. Data represent mean ± SEM. Bar in b is 91 µm and in e 35 µm. See Source Data file for numerical source data and unprocessed blots.