f4/80 antibody, anti-mouse, reafinity  (Miltenyi Biotec)

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

Journal: Scientific Reports

Article Title: Upregulation of IL-32γ in endothelial cells promotes macrophage recruitment and inflammatory progression in atherosclerosis

doi: 10.1038/s41598-026-40151-z

Figure Lengend Snippet: IL-32γ administration accelerates atherosclerosis progression in ApoE-/- mice through p38 MAPK signaling. ApoE−/− mice (6–8 weeks old, n = 6 per group) were fed a Western diet containing 0.2–1.25% cholesterol for 12 weeks and divided into three groups: ApoE−/− control, ApoE−/− + IL-32γ injection, and ApoE−/− + IL-32γ injection + SB202190 (p38 inhibitor). ( A ) ELISA measurements of IL-1β and TNF-α in serum. ( B ) Western blot analysis of p38 and p-p38 levels in macrophages/monocytes isolated from mouse blood using anti-F4/80 MicroBeads. ( C ) Oil Red O staining of whole aortas with quantification of atherosclerotic plaque area. ( D ) H&E staining of aortic sections with quantification of plaque lesion area. ( E ) Immunohistochemical staining for CD68 with quantification of macrophage infiltration in atherosclerotic plaques. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001.

Article Snippet: Monocytes/macrophages were then purified using anti-F4/80 MicroBeads UltraPure (Miltenyi Biotec, Cat# 130-110-443) according to the manufacturer’s protocol.

Techniques: Western Blot, Control, Injection, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Immunohistochemical staining