anti human mouse vegfa neutralizing antibody  (MedChemExpress)

Journal: Nature Communications

Article Title: Hypoxia-driven remodeling of SELENOP + macrophages shapes T cell dynamics and promotes ovarian cancer metastasis

doi: 10.1038/s41467-025-67859-2

Figure Lengend Snippet: a Dot plot of the Spearman correlation coefficients between distinct tumor MPs and the proportion of macrophage clusters (only showing p value < 0.05). p value are calculated by the two-sided Spearman correlation test with Benjamini–Hochberg adjustment. b Representative spatial co-localizations of SPP1 + macrophage with malignant epithelial cells with MP6 signature in our spatial transcriptomics cohort. c Heatmap showing the importances among all cell types signature scores in spots of spatial transcriptomics data. mEpi malignant epithelial cells. d Bubble heat map showing the mean interaction strength for selected ligand–receptor pairs between malignant epithelial cells and SELENOP + macrophages (malignant epithelial cells providing ligands) across groups. H_VS_L: the discrepancy in the interaction strength between malignant cells with high or low MP6 signature scores and SELENOP ⁺ macrophages. e Western blot images and quantifications of VEGFA in cell lines under normoxic or hypoxic conditions for 24, 48, and 72 h. f SELENOP in THP-1 or BMDMs after cocultured with conditioned medium (CM) from cell lines under normoxic or hypoxic conditions for 48 h, with or without VEGFA antibody (VEGFA Ab). Western blot images and quantifications of SELENOP after knocking down EPHB2 by CRISPR-Cas9 in THP-1 ( g ) or by shRNA in BMDMs ( i ) followed by rhVEGAF or rmVEGFA treatment. h ELISA showing SELENOP level in the culture supernatants in ( g ). For g – i , sgNC, empty vector for negative control of sgRNA; sgEPHB2, EPHB2 deletion mediated by CRISPR/Cas9; rh(m)VEGAF, recombinant human (mouse) VEGFA protein. shNC, transfected with negative control shRNA; KD1-Ephb2, Ephb2 knockdown by sh Ephb2 -1; KD2-Ephb2, Ephb 2 knockdown by sh Ephb 2-2. Western blot images and quantifications of VEGFA in PDOs under normoxic or hypoxic conditions for 72 h and 96 h ( j ). SELENOP in MDMs after cultured with CM from PDOs under normoxic or hypoxic conditions (96 h) ( k ). MDMs human monocyte-derived macrophages. l Representative images of PDOs under normoxic (upper) or hypoxic conditions (lower). For f , k , & denotes coculture. For j – l , PDOs patient-derived organoid, MDMs human monocyte-derived macrophages. For a , d , n = 17 scRNA-seq cohort solid site samples, biological replicates. For b , c , n = 24 spatial RNA cohort samples, biological replicates. For e – k , data represent the mean ± SD. For e , j , k , two-sided unpaired Student’s t test, n = 3, biological replicates ( e ) or technical replicates ( j , k ). For f – i , one-way ANOVA with Bonferroni post hoc test, n = 3, biological replicates. Source data are provided as a Source Data file.

Article Snippet: THP-1 cells and BMDMs were incubated with conditioned medium for 48 h, and rescue assays were subsequently conducted by supplementing the cultures with 50 μg/mL anti-human/mouse VEGFA neutralizing antibody (Cat#HY-P9906, MedChemExpress; Cat# 512810, BioLegend).

Techniques: Western Blot, CRISPR, shRNA, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Negative Control, Recombinant, Transfection, Knockdown, Cell Culture, Derivative Assay

Journal: Nature Communications

Article Title: Hypoxia-driven remodeling of SELENOP + macrophages shapes T cell dynamics and promotes ovarian cancer metastasis

doi: 10.1038/s41467-025-67859-2

Figure Lengend Snippet: a Schematic diagram of the protocol for in vivo experiments. C57BL/6J (B6) mice inoculated with ID8-luciferase cells are treated with 2.5 mg/kg IgG (control mouse; n = 9) or 2.5 mg/kg anti-VEGFA antibody (anti-VEGFA treatment mouse; n = 9). Created in BioRender. Song, X. (2025) https://BioRender.com/1zggfuh . Representative images ( b ) and quantifications ( c ) of C57BL/6J (B6) mice detected at 7 d (left), 14 d (middle) and 49 d (right) by bioluminescence imaging after inoculation with ID8-luciferase cells. 49 d after inoculation with ID8-luciferase cells, mice in both groups were euthanized ( n = 9 mice per group). Representative images showing characteristic ( d ) and quantifications ( e ) of peritoneal metastases (black arrows) in mice treated with control IgG (upper), and the absence of peritoneal metastases in mice treated with anti-VEGFA antibody (lower). Representative images ( f ) and quantification of tumor volumes ( g ) of primary tumors of mice treated with control IgG or anti-VEGFA antibody. Bars, 10 mm. Fresh primary tumor and peritoneal metastasis tissues were digested and stained for flow cytometry analysis. Shown is the proportion of F4/80 + CD11b + SELENOP + macrophages ( h ), CD3 + CD8 + GZMB + T cells ( i ), and CD3 + CD8 + PD-1 + T cells (j) across different sites of each group. Pri. Tumor, primary tumor of orthotopic ovarian cancer mouse model. Met.Per, peritoneal metastasis of orthotopic ovarian cancer mouse model. k Representative immunofluorescence staining showing co-localization of F4/80 (red), SELENOP (green), CD8A (magenta), GZMH (cyan) and DAPI (blue) in C57BL/6 J (B6) mice samples. l Blood serum SELENOP concentrations of mice treated with IgG or anti-VEGFA antibody. For c – j , l , n = 9 for each group, biological replicates, control IgG, control mouse group; anti-VEGFA, anti-VEGFA treatment mouse group. For c , e , g , h – j , l , data represent the mean ± SD, points represent individual samples. p values: two-sided unpaired Student’s t test for two groups, multiple t -test for multiple groups. ns not significant. Source data are provided as a Source Data file.

Article Snippet: THP-1 cells and BMDMs were incubated with conditioned medium for 48 h, and rescue assays were subsequently conducted by supplementing the cultures with 50 μg/mL anti-human/mouse VEGFA neutralizing antibody (Cat#HY-P9906, MedChemExpress; Cat# 512810, BioLegend).

Techniques: In Vivo, Luciferase, Control, Imaging, Staining, Flow Cytometry, Immunofluorescence

Journal: Nature Communications

Article Title: Hypoxia-driven remodeling of SELENOP + macrophages shapes T cell dynamics and promotes ovarian cancer metastasis

doi: 10.1038/s41467-025-67859-2

Figure Lengend Snippet: Schematic representation of the spatiotemporal heterogeneity of TME in cell type infiltration in the solid tumor sites of HGSOC, profiled by malignant epithelial cells, macrophages, and CD8 + T cells. Sketch map showing the proposed interaction model for the role of the malignant cells-macrophages-CD8 + T cells axis in the immunosuppressive TME formation. Under hypoxic conditions, malignant epithelial cells derived VEGFA amplification, leading to macrophages reprogramming, especially the decreased expression of SELENOP, followed by impaired cytotoxic function of precursor exhausted CD8 + T cells, associated with HGSOC metastasis. Created in BioRender. Song, X. (2025) https://BioRender.com/zm3gt6g .

Article Snippet: THP-1 cells and BMDMs were incubated with conditioned medium for 48 h, and rescue assays were subsequently conducted by supplementing the cultures with 50 μg/mL anti-human/mouse VEGFA neutralizing antibody (Cat#HY-P9906, MedChemExpress; Cat# 512810, BioLegend).

Techniques: Derivative Assay, Amplification, Expressing