Fig. 4: TSP1-KO alleviates glioma immunosuppression.

Enrichment plots and volcano plots summarizing GSEA with the gene sets Interferon Signaling (Reactome) (a) and TNF-mediated Signaling Pathway (GO:BP) b. Negative normalized enrichment scores (NES) indicate downregulation in WT tumors vs. KO. In volcano plots, gene set members are highlighted, leading-edge genes are labeled, and genes exceeding log₂FC or adjusted p value thresholds are shown at the edges. c Flow cytometry of brain-infiltrating leukocytes (BILs) using a tumor-associated macrophage (TAM)-related marker panel in mice with SB28-TSP1-WT or KO tumors. Representative contour plots show CD86+CD206− “pro-inflammatory” and CD86−CD206+ “anti-inflammatory” populations within CD45+CD11b+F4/80+ TAMs. Box plot shows the ratio of CD86+/CD206− to CD86−/CD206+ populations (n = 5 mice per group; p = 0.01; mean, 0.58 [WT] vs. 1.34 [KO]). Values in contour plots are percentages of gated populations. The box plot shows the median (center line), interquartile range (box limits), and minimum and maximum values (whiskers). d Histogram and bar plot summarizing the suppressive effect of TAMs on CD8+ T-cell proliferation. CD11b+ BILs were isolated from mice with SB28-TSP1-WT (n = 5) or KO (n = 4) tumors and co-cultured for 72 h with carboxyfluorescein succinimidyl ester (CFSE)-labeled T-cells isolated from a non-tumor-bearing mouse. The histograms show the peak distributions of CFSE signals in positive (PC) and negative controls (NC) and representative samples from WT and KO groups. The bar plot displays the percentages of proliferating T-cells. e Flow cytometry of BILs from mice with SB28-TSP1-WT or KO tumors, using a T-cell-related marker panel. Representative contour plots distinguish CD8+ and CD4+ populations within CD45+CD3+ T-cells. The bar plot shows the percentages of CD8+ T-cells identified in each sample (n = 4 mice per group; p = 0.01; mean: 21.4% [WT] vs. 71.5% [KO]). p values were calculated using the two-sided Welch’s unpaired t-test (c–e). For details on statistical tests used in (a) and (b), see “Methods”. In bar plots, data are mean ± s.e.m. d, e. Source data are provided as a Source Data file.