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. 2018 Aug 21;9:2862. doi: 10.1038/s41467-018-05099-3

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© The Author(s) 2018

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. 6 — Non-degraded NG2 in Lgl1 cKO OPC is recycled to the membrane. a Representative co-immunostaining for NG2-EC (red) and Rab11 (green) 45 min after incubation in differentiation medium. Note the higher colocalization of NG2-EC with Rab11 in Lgl1 cKO OPC (yellow arrowheads) compared to Lgl1 WT cells (red arrowhead). Scale bar: 5 μm. b–d Fluorescent immunostaining pictures illustrating colocalization of remaining NG2 (blue) with (b) ARF6 (red), (c) Rab4 (red), and (d) EEA1 (green) in Lgl1 cKO cells 24 hours after incubation in differentiation conditions. Scale bars: 10 μm. e, f Frames illustrating TIRF time lapse of Lgl1 cKO and WT cells 4 hours after incubation in differentiation conditions. Pictures were taken each 20 min. Examples of recycling labeled NG2 puncta (white) are highlighted in dashed circle. NG2 recycling persists in Lgl1 cKO cells up to time point 01:20 whereas it is insignificant in WT cells. Time indicated as hours:minutes. Scale bar: 10 μm. g Graph representing quantification of the corrected total cell fluorescence reflecting NG2 recycling in Lgl1 cKO cells (blue circles) and in Lgl1 WT cells (red lozenges). Data are depicted as mean ± s.e.m., n = 8 cells for each genotype, three independent experiments (***p < 0.01, two-way ANOVA)