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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. 3 — a Postnatal Rab35 expression in sciatic nerves. Represenative of two independent experiments. b Reduced Rab35 levels in sciatic nerve lysates from Rab35 cKO^SC mice at P30 compared to controls (Ctrl) set to 1. One-sample two-sided t-test, n = 5 pools of nerves per genotype in three independent experiments, p = 0.0001, t = 15.52, df = 4. Residual Rab35 is from fibroblasts and axons. c Semithin section analysis of control and Rab35 cKO^SC sciatic nerves at P20, P30, and P90. Red asterisks, examples of tomacula or increased myelin thickness. Yellow arrowheads, myelin outfoldings. Green arrowheads, myelin degenerations. Representative images for quantifications in f. d g-ratio at P20: Rab35 cKO^SC, 0.7 ± 0.005, 404 fibers; controls (Rab35^flox/flox and Rab35^flox/+), 0.7 ± 0.0024, 432 fibers, n = 5 mice/genotype, p = 0.69. e g-ratio at P90: Rab35 cKO^SC, 0.688 ± 0.002, 1162 fibers; controls (Rab35^flox/+), 0.682 ± , 0.002, 1499 fibers, n = 4 mice, p = 0.057. Fibers >5 µm in diameter: Rab35 cKO^SC, 0.73271374 ± 0.005, 130 fibers and control (Rab35^flox/+), 0.700473046 ± 0.0045, 446 fibers, n = 4 mice, p = 0.0286. d, e Two-tailed nonparametric Mann–Whitney test. f Myelinated fibers with aberrant myelin (in percentage of total): Rab35 cKO^SC at P20, n = 5 mice, percentage of Myelin degeneration: 3.053 ± 0.272. Tomacula: 3.684 ± 0.164. Myelin outfoldings: 2.829 ± 0.313; at P30, n = 4 mice, Myelin degeneration: 3.348 ± 0.221. Tomacula: 5.109 ± 0.508. Myelin outfoldings: 2.548 ± 0.136; at P70, n = 7 mice. Myelin degeneration: 5.390 ± 0.472. Tomacula: 6.916 ± 0.389. Myelin outfoldings: 9.336 ± 1.008, and at P90, n = 4 mice. Myelin degeneration: 6.993 ± 0.572; tomacula: 8.553 ± 0.676. Myelin outfoldings: 11.619 ± 0.588. g Representative images of ultrastructural analysis of control and Rab35 cKO^SC sciatic nerves (middle, right) at P90 quantified in f. Red asterisk, tomacula. Yellow arrowheads, myelin outfoldings. Green arrowhead, example of myelin degeneration. Mean ± SEM. Bar in c for P20, P30, and P90 semithin sections is 20 µm. Bar in g is 5 µm and 3.5 µm (for the outfolding and tomacula details). Numerical source data and unprocessed blots are reported in the Source Data file.