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. 2024 Apr 27;15:3599. doi: 10.1038/s41467-024-48010-z

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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. 8 — a Traced PVs in VTSs generated from CTRL, VEGF-A, or PDGF-B OE ZR75-1 tumor cells. PV segments are color-coded according to mean diameter. b Percentage of CD31⁺-volume of total VTS volume in ZR75-1 VTSs. CTRL and PDGF-OE: n = 4; VEGF-OE: n = 3. c Distribution of PV-segment mean diameters in ZR75-1 VTSs. CTRL and PDGF-OE: n = 4; VEGF-OE: n = 3. d Position of CD11b⁺-macrophages in CTRL, VEGF-A, or PDGF-B OE ZR75-1 VTSs. Frontal cut through VTS center in the xy-plane, 50 µm depth. e Density of CD11b⁺-macrophages in ZR75-1 VTSs. CTRL and PDGF-OE: n = 4; VEGF-OE: n = 3. f Distribution of minimal distance of CD11b⁺-macrophages to the VTS surface in ZR75-1 VTSs. CTRL and PDGF-OE: n = 4; VEGF-OE: n = 3. g Schematic view of the generation of Snai1/Twist1 OE MDA-MB-231 tumor cells and fibroblasts to produce genetically modified VTSs. h Principal component analysis of effects from Snai1 or Twist1 OE in MDA-MB-231 tumor cells or NHDFs. Analysis based on a 49-parameter descriptive data set. i Distribution of PV-segment diameters within VTSs generated from CTRL, Snai1, or Twist1 OE MDA-MB-231 cells. Average of 3 VTSs/group. j Distribution of PV-segment diameters within VTSs generated from CTRL, Snai1, or Twist1 OE NHDF. Average of 3 VTSs/group. k Traced PVs in VTSs generated from CTRL or Snai1 OE MDA-MB-231. Color-coded according to PV mean diameter. l Average distance of tumor cells to nearest pseudovessel in VTSs generated from CTRL, Snai1, or Twist1 OE NHDF. m PV networks (red) within a VTS (surface: transparent gray) generated from MDA-MB-231-GFP and NHDF-DsRed. Some PVs are located close to or on the surface of the VTS (white arrowheads). n Heatmap, displaying distances from the nearest PV (red) in a VTS generated from MDA-MB-231-GFP and NHDF-DsRed. The VTS’s center is almost void of PVs. o PV networks (red) within a VTS (surface: transparent gray) generated from MDA-MB-231-GFP and NHDF-Twist1. The outermost layer of PVs is substantially retracted from the surface of the VTS (yellow arrowheads). p Heatmap, displaying distances from the nearest PV (red) in a VTS generated from MDA-MB-231-GFP and NHDF-Twist1. PVs also pervade the center of the VTS. 3D grid spacing: 50 µm, scale bars: 100 µm error bars: ± SEM, analyzed with unpaired two-tailed t-test, n = 3, individual biological samples if not otherwise stated. Source data are provided as a Source Data file. OE overexpression.