mouse monoclonal antibody  (R&D Systems)

Journal: Journal of Translational Medicine

Article Title: Triple-remodeling of tumor microenvironment through hyaluronidase-assisted folate-targeted lipid nanoparticle-mediated siVEGF/siPD-L1 for enhanced tumor immunotherapy

doi: 10.1186/s12967-026-07697-y

Figure Lengend Snippet: In vitro evaluation of lipid nanoparticles. The images of K7 cells 4 h after uptake of Cy5-DNA ( A ) or FAM-siRNA ( B ) loaded by LNP or folate targeted FA-LNP. Scale bar = 20 μm. ( C ) Cell viability of FA-LNPs and FA-LNPvp in K7 cells after incubation for 48 h. ( D ) Quantitative experiment via flow cytometry after 48 h of FA-LNP/pEGFP transfection following 24 h of folate blockade. ( E ) Observation of GFP expression under fluorescence microscopy after 48 h of FA-LNP/pEGFP transfection following 24 h of folate blockade. Scale bar = 100 μm. ( F ) Detection of VEGF mRNA expression by RT-qPCR after 24 h of FA-LNPvp transfection in K7 cells. ( G ) Verification of PD-L1 protein expression on the surface of K7 cells by flow cytometry after 24 h of FA-LNPvp transfection. Results were expressed as means ± SD ( n = 3). * p < 0.05, **** p < 0.0001, ns = no significance

Article Snippet: An anti-PD-L1 recombinant monoclonal antibody was purchased from BioXcell, Inc. (West Lebanon, NH, USA).

Techniques: In Vitro, Incubation, Flow Cytometry, Transfection, Expressing, Fluorescence, Microscopy, Quantitative RT-PCR

Journal: Journal of Translational Medicine

Article Title: Triple-remodeling of tumor microenvironment through hyaluronidase-assisted folate-targeted lipid nanoparticle-mediated siVEGF/siPD-L1 for enhanced tumor immunotherapy

doi: 10.1186/s12967-026-07697-y

Figure Lengend Snippet: Combined anti-tumor mechanism of anti-angiogenic and immune checkpoint blockade therapy utilizing gene therapy. ( A ) The expression levels of VEGF mRNA in tumor tissues assessed using RT-qPCR. ( B ) Immunofluorescence labeling with CD31 positive neovascular growth within tumor tissues. Scale bar = 100 μm. ( C ) PD-L1 mRNA expression in tumor tissues quantified through RT-qPCR analysis. ( D ) Immunofluorescence labeling targeting PD-L1 within tumor tissues. Scale bar = 20 μm. Regarding the proportions of diverse immune cell subsets within tumor tissues: ( E ) CD45 + lymphocytes, ( F ) CD3 + CD8 + T cells, ( G ) PD-1 + Tim3 + CD8 + T cells within CD8 + T cell population, and ( H ) PD-1 + LAG3 + CD8 + T cells within CD8 + T cells. Results were expressed as means ± SD ( n = 6). ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = no significance

Article Snippet: An anti-PD-L1 recombinant monoclonal antibody was purchased from BioXcell, Inc. (West Lebanon, NH, USA).

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Labeling

Journal: Journal of Translational Medicine

Article Title: Triple-remodeling of tumor microenvironment through hyaluronidase-assisted folate-targeted lipid nanoparticle-mediated siVEGF/siPD-L1 for enhanced tumor immunotherapy

doi: 10.1186/s12967-026-07697-y

Figure Lengend Snippet: Anti-tumor immune mechanism study of FA-LNPs and FA-LNPvp. ( A ) H&E and immunofluorescence analyses of tumor tissue sections, H&E staining, cell proliferation marker Ki67, CD4⁺ T cells (red), and CD8⁺ T cells (green). Scale bars = 100 μm for H&E, 20 μm for Ki67 and CD4/CD8 immunofluorescence. Gene expression analysis in tumor tissues by RT-qPCR, ( B ) VEGF-A mRNA, ( C ) PD-L1 mRNA, and ( D ) HAase mRNA levels. Flow cytometric quantification of tumor-infiltrating immune cells, ( E ) CD3⁺CD4⁺ T cells, ( F ) CD3⁺CD8⁺ T cells, ( G ) CD11b⁺CD80⁺F4/80⁺ M1 macrophages, and ( H ) CD11b⁺Gr1⁺ M2 macrophages. Serum cytokine concentrations by ELISA, ( I ) IL-6, ( J ) TNF-α, and (K) IFN-γ levels in mouse serum. Results were expressed as means ± SD ( n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: An anti-PD-L1 recombinant monoclonal antibody was purchased from BioXcell, Inc. (West Lebanon, NH, USA).

Techniques: Immunofluorescence, Staining, Marker, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay