mouse vegf duoset elisa  (Bio-Techne corporation)

Journal: Journal of Extracellular Vesicles

Article Title: Mouse Tumor Tissue‐Derived Extracellular Vesicles Induce Angiogenesis Through VEGF Production From Macrophages

doi: 10.1002/jev2.70138

Figure Lengend Snippet: Tumor tEVs induce VEGF production from macrophages. (A) Overall scheme to investigate tumor tEV‐mediated VEGF production from macrophages. (B), (C) C57BL/6 mice were subcutaneously introduced with Matrigel (0.5 mL) containing PBS or tumor tEVs (10 µg). After 7 days, Matrigel was extracted, and paraffin sections of the Matrigel were immunostained with anti‐VEGF (in green) as well as anti‐F4/80 (in red) antibodies, and counter‐stained with Hoechst (in blue). (B) Images of paraffin sections stained for F4/80, VEGF, and Hoechst are shown. Scale bars represent 100 µm. (C) Fluorescence intensities of F4/80 staining were measured in VEGF‐negative or ‐positive areas within section images ( n = 6). (D) C57BL/6 mice were subcutaneously introduced with Matrigel (0.5 mL) containing PBS or tumor tEVs (10 µg). After 7 days, total RNA was isolated from Matrigel, and the level of VEGF mRNA was analyzed using real‐time RT‐PCR ( n = 3). Values represent the ratios of VEGF mRNA/GAPDH mRNA normalized to those in Matrigel containing PBS. (E) VEGF protein level in the conditioned media of peritoneal macrophages treated with a varying concentration of tumor tEVs for 24 h was measured by ELISA ( n = 4). Data are presented as mean ± SD. ** p < 0.01; and *** p < 0.001, calculated by two‐way ANOVA (C), unpaired Student's t ‐test (D), and one‐way ANOVA (E). Bonferroni correction for multiple comparisons was also performed for one‐ and two‐way ANOVA.

Article Snippet: Quantification of mouse VEGF protein was performed using DuoSet ELISA kit (R&D Systems) according to the manufacturer's protocol.

Techniques: Staining, Fluorescence, Isolation, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay