Journal: Cell Reports Medicine

Article Title: Targeting the CD40 costimulatory receptor to improve virotherapy efficacy in diffuse midline gliomas

doi: 10.1016/j.xcrm.2025.102204

Figure Lengend Snippet: The combination enhances a proinflammatory phenotype of the tumor myeloid compartment, with cDC1 playing a pivotal role in the anti-tumor response (A) Numbers of microglia, macrophages, and monocytes present per mg of tumor and assessed by flow cytometry in XFM tumors 6 days after being treated with the indicated conditions. (B and C) Representative images and analysis of the number of CD11c-positive cells (dendritic cells) assessed by immunofluorescence staining of XFM tumors at day six post-treatment. Scale bar: 100 μm. (D and E) Number and percentage of dendritic cells (DCs) from the total immune tumor infiltrate. (F) The concentration of the indicated proinflammatory chemokines and cytokines in the tumor microenvironment 6 days after treatment was assessed by LEGENDplex. (G) Volcano plot showing the differentially expressed genes (assessed by RNA-seq) in XFM tumors treated with the combination compared to IgG control at day 6. (H) Gene set enrichment analysis of pathways involved in myeloid cell activation and differentiation present in Delta-24-RGD+anti-CD40-treated tumors compared to IgG control and anti-CD40 alone. (I) Mean of fluorescence intensity (MFI) of MHC-II on microglia and tumor macrophages was measured by flow cytometry 6 days after treatment. (J) Number of DCs expressing high levels of MHC-II molecules per tumor mg. (K) Number of tumor DCs expressing CD40 and CD86 per tumor mg. (L) MFI of PDL1 present on the membrane of tumor DCs at day 6 post-treatment. (M) Representative example of flow cytometry plots showing the mCherry signal in the indicated populations infiltrating the NP53-mCherry orthotopic DMG model. (N) Quantification of the mCherry-positive cells from (E) in the tumor niche 24 h after the indicated treatments. (O) Survival curve of Batf3ko mice bearing UC-BL6-C7 tumor and treated with either the combination or an IgG control. (P) Representative H&E images of two brains treated with the IgG control and the combination that were collected at the endpoint. Error bars represent mean ± SEM. One-way ANOVA and the Kruskal-Wallis test were used for statistical analyses from (A)–(H) data. n = 3–10 mice per group. Log rank test was used for statistical analysis comparing the combination with IgG control-treated Batf3ko mice in the survival experiments. n = 4 mice per group. (∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001).

Article Snippet: Anti-mouse CD11c , CellSignalling , Cat#97585; RRID: AB_2800282.

Techniques: Flow Cytometry, Immunofluorescence, Staining, Concentration Assay, RNA Sequencing, Control, Activation Assay, Fluorescence, Expressing, Membrane

Journal: Cell Death & Disease

Article Title: The P-selectin and PSGL-1 axis accelerates atherosclerosis via activation of dendritic cells by the TLR4 signaling pathway

doi: 10.1038/s41419-019-1736-5

Figure Lengend Snippet: Detection of peripheral blood dendritic cells (DCs) by standardized 3-color flow cytometry. Peripheral blood samples were collected and the cells were stained with human phycoerythrin (PE) anti-CD11c, PE anti-CD123, peridinin chlorophyll protein anti-HLA-DR, and fluorescent isothiocyanate (FITC) lineage cocktail 1 (lin1). (Aa): Events excluding debris and dead cells (P1); (Ab): cells were gated on region P1, a dot blot of lineage marker ( x- axis) vs. HLA-DR ( y- axis) was used to define the region of Lin − cells (P2); (Ac) pDC was defined as Lin1 − HLA−DR + CD123 + (P4); (Ad) mDC was defined as Lin1 − HLA-DR + CD11c + (P5). Flow cytometric analysis of the percentage and absolute numbers of mDCs (Ba and Bb), percentages and numbers of pDCs (Bc), and the mDC/pDC ratio (Bd) in control subjects ( n = 34) and STEMI patients ( n = 34) on admission. ELISA analysis of circulating P-selectin (Ca) and flow cytometric analysis of the percentage and absolute numbers of mDCs (Cb and Cc), and the mDC/pDC ratio (Cd) of STEMI patients on admission and at 5–7 days after PCI ( n = 34). The circulating mDC/pDC ratio was inversely correlated with serum P-selectin levels in STEMI patients on admission (Da), 5–7 days after successful PCI (Db), and in total (Dc), while no such correlation was found in controls (Dd). mDC: myeloid dendritic cell; pDC plasmacytoid dendritic cell

Article Snippet: DCs in the sections were examined using an Alexa Fluor ® 647-conjugated anti-CD11c Ab (1:50; Novus Biologicals LLC, Littleton, CO, USA).

Techniques: Flow Cytometry, Staining, Dot Blot, Marker, Control, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: The P-selectin and PSGL-1 axis accelerates atherosclerosis via activation of dendritic cells by the TLR4 signaling pathway

doi: 10.1038/s41419-019-1736-5

Figure Lengend Snippet: ApoE −/− , ApoE −/− P −/− , and ApoE −/− PSGL-1 −/− mice were fed a western diet for 12 weeks. a Representative flow cytometric dot plots showing the percentage of CD11c + DCs expressing the DC maturation marker CD86 or MHC- II in spleen tissue. b ELISA analysis of the indicated cytokine levels in serum. WT and PSGL1 −/− mice were subcutaneously infused with P-selectin, NS, or LPS for 1 day. c , e Representative flow cytometric dot plots showed the percentage of CD11c + DCs expressing the DC maturation marker CD86 or MHC-II in spleen tissue. d , f qRT-PCR analysis of the indicated cytokine mRNA levels in spleen tissue. Data are expressed as the mean ± SD ( n ≥ 5). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: DCs in the sections were examined using an Alexa Fluor ® 647-conjugated anti-CD11c Ab (1:50; Novus Biologicals LLC, Littleton, CO, USA).

Techniques: Western Blot, Expressing, Marker, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: The P-selectin and PSGL-1 axis accelerates atherosclerosis via activation of dendritic cells by the TLR4 signaling pathway

doi: 10.1038/s41419-019-1736-5

Figure Lengend Snippet: ApoE −/− , ApoE −/− P −/− , and ApoE −/− PSGL-1 −/− mice were fed a western diet for 12 weeks. a Representative images of Oil red O-stained aortae after en face preparation and quantification of lipid content (lipid content/vascular area) in three different groups. b Representative histological analysis results of the aortic sinus stained with H&E, Movat, and Masson’s trichrome stain, and quantification of the plaque area (plaque area/lumen area), necrotic area (necrotic area/ plaque area), and collagen content (collagen content/plaque area) in aortic sinus. Scale bar: 200 μm. c Representative immunofluorescence staining of the aortic sinus of lesions with Alexa Fluor ® 647-conjugated anti-CD11c Ab. Scale bars: 200 and 50 μm, respectively. Data are expressed as the mean ± SD ( n = 8). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: DCs in the sections were examined using an Alexa Fluor ® 647-conjugated anti-CD11c Ab (1:50; Novus Biologicals LLC, Littleton, CO, USA).

Techniques: Western Blot, Staining, Immunofluorescence

Journal: iScience

Article Title: Macrophages treated with interferons induce different responses in lymphocytes via extracellular vesicles

doi: 10.1016/j.isci.2024.109960

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti CD11c (clone REAA618) Conjugated APC , Miltenyi , Cat#130-114-110.

Techniques: Recombinant, Protein Extraction, Protease Inhibitor, Pore Size, Electrophoresis, Staining, Software, Cytometry