rat anti mouse monoclonal f4 80  (Bio-Rad)

Journal: Cancer Cell

Article Title: IGSF11-VISTA is a critical and targetable immune checkpoint axis in diffuse midline glioma

doi: 10.1016/j.ccell.2025.12.020

Figure Lengend Snippet: DMG tumor-immune spatial organization and the IGSF11-VISTA pathway are preserved in mice (A) Schematic representation of the in-utero electroporation (IUE) mouse model dependent on DMG driver mutations. (B) Dot plot representation of the co-occurrence patterns in Visium ST data ( n = 4). Dot plots reflect estimated relative NMF weights normalized across components on a color and size scale. (C) Visium ST data depicting estimation of DMG cancer cell states abundance (color intensity) overlayed on top of the tissue image (left). Spatial representation of the co-occurrence patterns, shown as compartment enrichment (color intensity) with a dashed line delineating the tumor border (right). (D) Heatmap visualization of z-scaled expression scores (blue-to-red color gradient) for biological and immune processes across mouse healthy brain and AOO- and MES-enriched tumor areas ( n = 4). (E) UMAP representation of mouse microglia sub-populations ( n = 4). (F) Violin plot of MHC-I (purple) and MHC-II (pink) related gene signature expression across microglia sub-populations ( n = 4). ∗∗∗∗ p < 0.0001, Mann-Whitney U test. (G) Dot plot (left) of tumor and immune cell co-occurrence patterns in ST datasets, segregating between homeostatic and inflammatory microglia ( n = 4). Dot plots reflect estimated relative NMF weights normalized across components on a color and size scale. Spatial maps (right) of compartment enrichment (color intensity) for homeostatic and inflammatory microglia. Dashed line represents the AOO- (yellow) and MES-enriched (orange) areas. (H) Relative expression (color intensity) of Vsir across immune populations ( n = 4). Datapoint size indicates the percentage of expressing cells. (I) Relative expression (color intensity) of immune checkpoints in mouse DMG tumor cells ( n = 4). Datapoint size indicates the percentage of expressing cells. (J) Relative expression (color intensity) of Igsf11 across mouse DMG tumor populations ( n = 4). Datapoint size indicates the percentage of expressing cells. (K) 2D overview image (left) and zoom in (right) on the area outlined by a dashed square (i) of a mouse DMG tumor-bearing brain. Tissue slice stained for DAPI (gray), tumor-eGFP (green), IGSF11 (red), IBA1 (magenta), VISTA (yellow), F4/80 (orange), and CD3 (cyan). Scale bars, 1mm (overview - left) and 50μm (zoom - right). (L and M) Frequency of Igsf11 + (L) or Vista + (M) cells in the tumor compared to the healthy brain region ( n = 6). Boxplots show the interquartile range with the median indicated, and whiskers represent 1.5x the interquartile range. ∗∗ p < 0.01, Wilcoxon test, ns = not significant.

Article Snippet: Human anti-F4/80 antibody , Miltenyi Biotec , Cat# 130-116-499; RRID: AB_2727574.

Techniques: In Utero, Electroporation, Expressing, MANN-WHITNEY, Staining

Journal: Cancer Cell

Article Title: IGSF11-VISTA is a critical and targetable immune checkpoint axis in diffuse midline glioma

doi: 10.1016/j.ccell.2025.12.020

Figure Lengend Snippet: Igsf11 signaling reinforces VISTA expression and maintains microglia in a homeostatic state (A) Quantification of survival study endpoint eGFP+ tumor area (left) in the pons of mice receiving Igsf11 WT ( n = 6) or OE ( n = 7) tumor cells. Bar graphs indicate the mean ± SEM. ∗ p < 0.05, t test. Representative stereo images (right) of the tumor signal (green) in the pontine region. Scale bars, 5mm. (B) Kaplan-Meier survival curves of immunocompetent C57Bl/6 mice allografted with WT ( n = 7), KD ( n = 10), or Igsf11 OE tumor cells ( n = 10). ∗∗∗∗ p < 0.0001, Log rank Mantel-Cox test. (C) Immune cell frequencies in Igsf11 WT (green) and Igsf11 OE (orange) DMG tumors (n = 5–6). Boxplots show the interquartile range with the median indicated, and whiskers represent 1.5x the interquartile range. ∗∗∗ p < 0.001, t test. (D–F) Frequency of Vista + (D), MHCII + (E), or CD206 + (F) microglia in Igsf11 WT (green) and OE (orange) DMG tumors (n = 5–6). Boxplots show the interquartile range with the median indicated, and whiskers represent 1.5x the interquartile range. ∗ p < 0.05, ∗∗∗ p < 0.001, t test. (G) 2D overview image of an Igsf11 WT and OE tumor tissue slice stained for DAPI (gray), eGFP-Tumor (green), Igsf11 (blue), VISTA (white), IBA1 (magenta), F4/80 (orange), MHCII (yellow), TREM2 (red), CD206 (green), SPP1 (pink), and GPNMB (cyan). Scale bars, 50μm.

Article Snippet: Human anti-F4/80 antibody , Miltenyi Biotec , Cat# 130-116-499; RRID: AB_2727574.

Techniques: Expressing, Staining