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. 2023 Feb 1;43:8. doi: 10.1186/s41232-022-00254-2

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© The Author(s) 2023

Open AccessThis 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. 3. — VEGF-A is secreted from NSPCs and enhances neurosphere formation under hypoxic condition. A Quantification of neurospheres from NSPCs by administration of VEGF-A (mean ± SEM, n = 3, ** p = 0.00754, * p = 0.0436, (^a p = 0.00377, ^b p = 0.0305, ^c p = 0.0218, ^d p = 0.0473 to the control in the case of one-sided test)). B Quantification of neurospheres from NSPCs by administration of 10 ng/ml of VEGF-A and 700 nM of VEGFR2 inhibitor, SU1498 (mean ± SEM, n = 3, ^a p = 0.00555, ^b p = 0.00695). The equivalent volume of DMSO which was used as solvent of SU1498 was administrated in the control. C Neurosphere formation from enriched E14.5 NSPCs in the presence of SU1498 (700 nM) under the normoxic condition. D VEGF-A gene expression in NSPCs isolated from E11.5 brain under the hypoxic condition was analyzed by semi-quantitative reverse transcription-PCR. The gel images were cropped and the full-length gel images were shown in Fig. S12A. E Measurement of VEGF-A in the E11.5 NSPC culture under the normoxic or the hypoxic condition by ELISA (mean ± SEM, n = 4, ** p = 0.002858). Each black dot indicates the VEGF-A values in each sample. F VEGF-A gene expression in NSPCs isolated from E14.5 brain under the hypoxic condition was analyzed by semi-quantitative reverse transcription-PCR. The gel images were cropped and the full-length gel images were shown in Fig. S12B, C. The gel image of VEGF-A was inverted horizontally. G Measurement of VEGF-A in the E14.5 NSPC culture under the normoxic or the hypoxic conditions by ELISA (mean ± SEM, n = 3, * p = 0.01448). H Quantification of neurospheres of NSPCs formed in the hypoxic neurosphere formation culture supplemented with or without SU1498 (mean ± SEM, n = 4, * p = 0.0482). I Quantification of neurospheres of NSPCs formed in the conventional normoxic neurosphere formation culture with the hypoxic NSPC CM, supplemented with or without SU1498 (mean ± SEM, n = 4, * p = 0.0107)