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. 2024 Oct 9;15:8570. doi: 10.1038/s41467-024-52444-w

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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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Fig. 2 — NRP1^+/+ and NRP1⁻^/− oligodendroglia were incubated for 3 days with or without 5µg/ml SEMA3A. This did not affect the number of NRP1^+/+ or NRP1^−/− PDGFRα + OPCs (A), two-tailed unpaired t test, p = 0.0798 and 0.7139 respectively, or the number of OLIG2+ oligodendroglia (B), two-tailed unpaired t test, p = 0.9394 and 0.2686 respectively, n = 5 for all conditions. C Oligodendroglia proliferation was assessed with KI67, and SEMA3A had no effect on proliferation of NRP1^+/+, n = 4, two-tailed unpaired t test, p = 0.8908 or NRP1⁻^/− oligodendroglia, n = 5, two-tailed unpaired t test, p = 0.4602. D Flow cytometry of live/dead cells using marker draq7 found SEMA3A did not affect cell survival, n = 3 for all conditions, two-tailed unpaired t test, NRP1^+/+ p = 0.2814, NRP1⁻^/− p = 0.8788. E A significant decrease in O4 + oligodendrocytes was observed when NRP1^+/+ differentiating cultures were incubated with SEMA3A, an effect not observed in NRP1^−/− oligodendrocytes. n = 5 for all conditions, two-tailed unpaired t test, NRP1^+/+ *p = 0.0330, NRP1⁻^/− p = 0.3706. F Representative images showing NRP1^+/+ and NRP1⁻^/− O4 + oligodendrocytes following exposure to SEMA3A, scale bar 100 μm (G) NRP1^+/+ cells were incubated with 5 µg/ml SEMA3A for 1 week. No difference in mature MBP + oligodendrocytes was observed between the treated and untreated groups, n = 4 for both conditions, two-tailed unpaired t test, p = 0.6112. H Representative images of MBP + NRP1^+/+ oligodendrocytes following one-week incubation with SEMA3A, scale bar 100 μm. I NRP1^+/+, and NRP1⁻^/− cells were placed in transwells with untreated base media or base media containing 5 µg/ml SEMA3A. Hoechst was used to quantify the total number of cells, and the number of migrated cells was used to calculate the % of migrated cells. SEMA3A reduced NRP1^+/+ cell migration by 33% but did not affect the migration of NRP1⁻^/− cells. n = 3 for all conditions, two-tailed unpaired t test, NRP1^+/+ ***p = 0.0002, NRP1⁻^/− p = 0.8382. All graphs plot means ± SEM. Each point represents a separate differentiation culture. Source data are provided in the Source Data file.