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. 2024 May 30;27(8):1555–1564. doi: 10.1038/s41593-024-01654-y

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 8 — (a) Immunohistochemistry performed in the cingulum of 1-month-old Nf1^WT and Nf1^OPC-iKO mice (tamoxifen injected at P28) revealed that Nf1^OPC-iKO mice had an increased percentage of new OPCs (the number of EdU⁺/Olig2⁺/PDGFRα⁺ cells divided by the number of EdU⁺/Olig2⁺ cells) and reduced percentage of new OLs (the number of EdU⁺/Olig2⁺/PDGFRα^- cells divided by the number of EdU⁺/Olig2⁺ cells) relative to Nf1^WT mice. N = 4 per group. ***, P = 0.0005. Welch’s test. (b) Nf1^WT and Nf1^OPC-iKO mice received tamoxifen at 13 weeks of age and pulsed with EdU in the drinking water for one week starting at 15 weeks of age. Immunohistochemistry performed at 16 weeks of age revealed that Nf1^OPC-iKO mice (N = 3) had an increased percentage of new OPCs (the number of EdU⁺/Olig2⁺/PDGFRα⁺ cells divided by the number of EdU⁺/Olig2⁺ cells) and reduced percentage of new OLs (the number of EdU⁺/Olig2⁺/PDGFRα^- cells divided by the number of EdU⁺/Olig2⁺ cells) relative to Nf1^WT mice (N = 6). Mann-Whitney test; *, P = 0.0238. (c) Transmission electron microscopy (TEM) analysis of myelin structure in the cingulum of 4-month-old Nf1^WT and Nf1^OPC-iKO mice revealed similar g-ratios (N = 4 per group) and densities of myelinated axons (N = 4 per group). Welch’s test. (d) Stereological analysis revealed similar cingulum volume between 4-month-old Nf1^WT (N = 5) and Nf1^OPC-iKO (N = 3) mice. Welch’s test. (e) TEM revealed similar g-ratios and density of myelinated axons in the corpus callosum of 4-month-old Nf1^WT (N = 5) and Nf1^OPC-iKO (N = 4) mice. Welch’s test. (f) TEM revealed similar g-ratios in the corpus callosum of 3-month-old WT and Nf1^+/- mice. N = 4 per group. Welch’s test. (g) Immunohistochemistry revealed a similar density of oligodendrocytes (ASPA⁺ cells) in the cingulum and corpus callosum of 6-month-old WT (N = 3) and Nf1^+/- (N = 4) mice. Welch’s test (cingulum). Mann-Whitney test (corpus callosum). (h) Stereological analysis revealed similar corpus callosum volume between WT and Nf1^+/- mice at 1-month and 3-month of age. N = 5 per group. Welch’s test. Data shown as mean ± SEM; each point = one mouse (a-h); two-sided; ns, not significant (P > 0.05). Scale bars, 2 µm.

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