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. 2019 Dec 17;10:5758. doi: 10.1038/s41467-019-13640-1

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© The Author(s) 2019

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. 4 — a, c Immunofluorescence analysis revealed decreased expression of vascular endothelial growth factor receptor (VEGFR) 1, VEGFR2, and increased expression of platelet-derived growth factor receptor-beta (PDGFR-β) in schwannomas post vaccination in case 1 (a) and case 2 (c). b, d Quantitative real-time PCR analysis showed decreased mean relative expression of vascular endothelial growth factor (VEGF)-A, VEGFR1, and VEGFR2 genes in schwannomas post vaccination in case 1 (b) and case 2 (d) (pre, n = 1 for cases 1 and 2; post, n = 1 for cases 1 and 2, respectively). e VEGF-A expression was lower after vaccination than before vaccination in cases 1 and 2. f Vessel diameter was smaller, and microvessel density was lower after vaccination in cases 1 and 2. g The numbers of Foxp3-positive cells and FOXP3 gene expression were lower in tumors after vaccination in cases 1 and 2 (pre, n = 1 for cases 1 and 2; post, n = 1 for cases 1 and 2, respectively). The black arrows indicate Foxp3-positive cells. h The total numbers of CD8-positive cells did not change substantially after vaccination. The numbers of CD8-positive cells in the perivascular area increased in case 2. i After vaccination, more cleaved caspase 3-positive cells were detected in tumor cells and endothelial cells in cases 1 and 2. In the panels showing VEGF-A and microvessel density, the original magnification was ×200 and the magnification bars = 100 μm. In all other panels, the original magnification was ×400 and the magnification bars = 100 μm. VEGFR vascular endothelial growth factor receptor, PDGFR-β platelet-derived growth factor receptor-beta, DAPI 4′,6-diamidino-2-phenylindole, VEGF vascular endothelial growth factor, MVD microvessel density.