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. 2021 Jun 23;12:3895. doi: 10.1038/s41467-021-23995-z

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© The Author(s) 2021

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. 3 — a Representative immunofluorescence appearance of CD34⁺ (red)/CD45⁺ (green) cells (arrows) in formalin-fixed, paraffin-embedded glioblastoma tissue sections from two patients (out of four analyzed, all with similar results). Nuclei were counterstained with DAPI (blue). Scale bars = 50 µm (overview) and 10 µm (insets). b Diagram describes the hierarchy of HSPC subsets analyzed in this study by flow cytometry. HSC Hematopoietic stem cell, MPP Multipotent progenitor, MLP Multi-lymphoid progenitor, CMP–MEP Common myeloid progenitor and megakaryocyte-erythroid progenitor, GMP Granulocyte–Monocyte Progenitor, B-NK B-NK progenitor. c Representative flow cytometry profiles of human bone marrow and glioblastoma tissue, gated for seven HSPC subsets. Cellular frequencies are highlighted in red. d Stacked barplot of seven HSPC subsets observed in glioblastoma (GBM, n = 7), non-glioblastoma tumor tissues (non-GBM, n = 5, Supplementary Data 4) and a healthy donor bone marrow sample by flow cytometry. e Representative colony morphologies from CFC assays of glioblastoma cell suspensions derived from a total of eight patients, and from bone marrow-derived mononuclear cells. Scale bars = 100 µm. f Barplot indicating number and types of colonies in CFC assays derived from bone marrow, glioblastoma (GBM, n = 8 patients) and non-glioblastoma (non-GBM, n = 6 patients) cell suspensions. CFU Colony forming unit, -GEMM Granulocyte, erythrocyte, monocyte, megakaryocyte, -GM, Granulocyte, monocyte, -E, Erythroid. g CXCL12 staining of the infiltration zone (upper panel) and the tumor (lower panel) of the same tissue section. Scale bars = 200 µm. One representative staining of eight shown). h Barplot showing percent CXCL12 staining intensity (1 = weakly positive, 2 = moderately positive, 3 = strongly positive) of tumor core and infiltration zones from glioblastoma (n = 16 samples from eight patients). Source data of (a), (d), (f), (g), and (h) are provided as a Source Data file.