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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. 10 — a, b Schwann cells from Rab35^flox/flox cKO or non-Cre^ER-Rab35^flox/flox mice (WT) mice treated with DMSO, VPS34-IN1, Cmp-19, Apilimod, or Rapamycin. a Images of DMSO- or inhibitor-treated Rab35 WT and cKO cells immunostained for p-S6 (green), total S6 (red). DAPI (blue), nuclei. b Quantification of representative data in a. Normalized (to DMSO-WT) p-S6/S6 ratio from n experiments. One sample two-tailed Student’s t-test (theoretical means of 1). DMSO: n = 6, ^*p = 0.0249, t = 3.168, df = 5; VPS34-IN1: n = 3, p = 0.2059, t = 1.848, df = 2; Cmp-19: n = 4, p = 0.5464, t = 0.6779, df = 3; Apilimod: n = 4, p = 0.7586, t = 0.3366, df = 3; Rapamycin: n = 3, p = 0.2241, t = 1.739, df = 2. c Images of differentiated DMSO- or inhibitor-treated Rab35 WT or cKO Schwann cells. PI(3)P (green), F-actin (red), DAPI (blue), and nuclei. d Quantification of representative data in c from n experiments. Normalized (to DMSO-WT set to 100%) PI(3)P intensity/cell area. One sample two-tailed student’s t-test (theoretical mean of 100). DMSO: n = 4, ^*p = 0.027, t = 4.058, df = 3; Apilimod: n = 3, p = 0.3237, t = 1.298, df = 2. e Images of inhibitor-treated Rab35 WT or cKO Schwann cells. P0 (green), F-actin (gray). f Quantification of representative data in e from n experiments. Intensity of P0/cell number (in percentage, DMSO-WT as 100%) (see Supplementary Fig. 8i). One sample two-tailed student’s t-test (theoretical means of 100). DMSO: n = 8, ^**p = 0.0014, t = 5.102, df = 7; VPS34-IN1: n = 5, p = 0.2093, t = 0.1.495, df = 4; Cmp-19: n = 6, p = 0.2510, t = 0.1.298, df = 5; Apilimod: n = 6, p = 0.3190, t = 0.7627, df = 5; Rapamycin: n = 4, p = 0.7541, t = 0. 3432, df = 3. Scale bars, 10 µm (a, c) or 50 µm (e). n.s. non-significant in all panels. Mean ± SEM. See Source Data file for numerical source data and unprocessed blots.