Fig. 3: Perivascular cells are slow-cycling.

a Left: Distribution of ki67-positive cells in the perivascular and parenchymal compartment in orthotopic S24 (n = 16 regions from three mice, two-tailed t-test) and P3xx (n = 12 regions from four mice, two-tailed t-test) xenograft tumors. Right: ki67 proliferation index (ki67 positive cells/all cells in the respective compartment) of S24 (n = 3 mice, two-tailed t-test) and P3xx tumor cells (n = 4 mice, two-tailed t-test) in the perivascular and the parenchymal compartment in xenograft tumors. b Exemplary immunofluorescence stainings of ki67 (yellow, proliferating cells), nestin (green, tumor cells), DAPI (blue, nuclei), and CD31 (red, endothelium) in S24 and P3xx xenografts. c Left: Quantification of the distribution of EdU-positive tumor cells (indicating cells in S-phase) after in vivo incorporation of EdU (4 h) in the parenchymal and perivascular compartment (S24 xenografts, n = 25 regions from 5 mice, two-tailed t-test; P3xx xenografts, n = 12 regions from 4 mice, two-tailed t-test). Right: EdU labeling index (EdU positive cells/all cells in the respective compartment) in the perivascular and the parenchymal compartment (S24 xenografts, n = 5 mice, two-tailed t-test; P3xx xenografts, n = 4 mice, two-tailed t-test). d Exemplary immunofluorescence staining of EdU (green, cells in S-phase), nestin (red, tumor cells), DAPI (blue, nuclei) and CD31 (yellow, endothelium) demonstrates that most proliferating cells are located in the parenchymal compartment (P3xx xenograft). Blood vessels are marked with arrowheads. e Mitotic index (number of mitotic events normalized to the distribution of cells in the perivascular and parenchymal compartment) of S24 and T269 GBMSCs (n = 16 regions in eight mice (S24), n = 10 regions in 4 mice (T269), two-tailed Mann Whitney tests). Right: Exemplary in vivo 2-PM of mitosis of a parenchymal S24 GBMSC (arrowhead). Cytoplasm (red), nucleus (green), blood vessels (blue). f Distribution of ki67 positive cells in the parenchymal and perivascular compartment in molecularly defined human glioma specimen (oligodendroglioma: n = 16 patients; astrocytoma: n = 19 patients; high grade astrocytoma: n = 15 patients; glioblastoma: n = 16 patients; one-way ANOVA (p = 1.36 × 10⁻¹³⁹), Tukey’s post hoc test for multiple comparisons). g Exemplary immunohistochemical stainings of ki67 in molecularly defined human glioma specimen. Arrowheads exemplarily mark blood vessels. Data in violin plots a, c, e, f are represented as median and quartiles. Data in columns a are represented as mean + SEM. *p < 0.05**, p < 0.01, ***p < 0.001. (mut: mutant; LOH: loss of heterozygosity; wt: wild-type). Source data are provided as a Source Data file.