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. 2020 Sep 24;10:15676. doi: 10.1038/s41598-020-72485-7

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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 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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HG induced alterations in SLIT-ROBO signalling pathway genes. (a) Western blot analysis revealed the decreased protein expression levels of SLIT1, ROBO2, SRGAP1 and CDC42 in hNPCs exposed to HG. (b) Quantitative analysis depicts significant decrease in SLIT1 (168 kDa), ROBO2 (172 kDa), SRGAP1 (124 kDa) and CDC42 (21 kDa) protein expression in hNPCs exposed to HG condition. Data is represented as Mean ± SE, n = 3, *p < 0.05, **p < 0.01, ***p < 0.001. Uncropped blot images are shown in Supplementary Fig. S2. Further, the decrease in the expression of ROBO2 (c, Red), SRGAP1 (d, Red) and CDC42 (e, Green) in hNPCs was confirmed by immunocytochemical analysis. Musashi1 (Green) and Nestin (Red) was used to mark neural stem cells while DAPI was used to stain the nuclei (scale bar = 10 μm). (f) Furthermore, the protein expression of SLIT1 (A,B, Green), ROBO2 (C,D, Red), SRGAP1 (E,F, Red) and CDC42 (G,H, Green) were localised in forebrain sections of mouse embryos from normal and diabetic pregnancy. The expression levels of SLIT1, ROBO2, SRGAP1 and CDC42 appear to be markedly reduced in forebrain sections of mouse embryos diabetic pregnancy, compared to that of embryos from normal pregnancy, scale bar = 200 μm. The third ventricle is marked as III.