human vegf duoset elisa kit  (R&D Systems)

Journal: Scientific Reports

Article Title: The heterogeneity of NOTCH1 to tumor immune infiltration in pan-cancer

doi: 10.1038/s41598-024-79883-1

Figure Lengend Snippet: The correlation of NOTCH1 with angiogenesis and angiopoietins. The KEGG pathways of NOTCH1 and its ligand protein of the GSEA gene set; Different colors respectively represent the enriched signal pathway, and the deeper the color gradient, the smaller the correlation of pvalue value (A) ; VEGF expression in the supernatant culture medium of three kind cells were detected by ELISA; * means p < 0.05 (B) ; The correlation between NOTCH1 and genes of angiogenesis; The correlation between NOTCH1 and genes of angiopoietin, p < 0.05 had significant correlation (C) .

Article Snippet: The VEGF content of cell culture supernatant was measured using human VEGF DuoSet ELISA kit (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: The Reduction of PSMB4 in T24 and J82 Bladder Cancer Cells Inhibits the Angiogenesis and Migration of Endothelial Cells

doi: 10.3390/ijms25105559

Figure Lengend Snippet: The effect of PSMB4 knockdown on HUVEC angiogenesis. ( A ) The conditioned medium of RT4, T24, and J82 cells transfected with siPSMB4 was collected. HUVECs were cultured with these conditioned media for 6 h. The tube formation ability was evaluated. Scale bar = 200 μm. ( B , C ) The length and branching of formed tubes were analyzed ( n = 5). ( D ) The VEGF protein content in the conditioned medium of RT4, T24, and J82 cells after treatment with siPSMB4 was measured with an ELISA kit ( n = 4). ( E ) The mRNA levels of VEGF-B in RT4, T24, and J82 cells after treatment with siPSMB4 for 72 h were measured via real-time PCR ( n = 4). ( F ) VEGF-B protein expression after siPSMB4 transfection for 72 h in RT4, T24, and J82 cells ( n = 7). Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the siRNA negative control group, determined by unpaired t-test with the Mann–Whitney test.

Article Snippet: The ELISA kit for VEGF was purchased from R&D Systems (DY293B-05, Minneapolis, MN, USA), and the concentration of VEGF in the culture medium was measured according to the manufacturer’s instructions.

Techniques: Knockdown, Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing, Negative Control, MANN-WHITNEY

Journal: Molecular oncology

Article Title: Spontaneous metastasis xenograft models link CD44 isoform 4 to angiogenesis, hypoxia, EMT and mitochondria-related pathways in colorectal cancer.

doi: 10.1002/1878-0261.13535

Figure Lengend Snippet: Fig. 6. Colony-forming capacity of and VEGF release from HT-29 spheres in normoxia vs. hypoxia upon CD44 kd. (A) Validation of the used HIF1a immunostaining protocol using HT-29 cells cultivated under normoxia or hypoxia (representative example is shown; n = 2), scale = 50 lm. (B) Flow cytometric analysis of pan-CD44 and CD44v expression on HT-29 cells recovered from 3D spheres cultured in nor- moxia (N) vs. hypoxia (H) (representative example is shown; n = 2). (C) 3D colony-forming ability of CD44 kd compared to control cells in N vs. H in Matrigel containing HA. *P = 0.037 in (C) refers to CD44 kd H vs. CD44 kd N (n = 5). (D) VEGF release per colony area from 3D control vs. CD44 kd HT-29 spheres grown in Matrigel containing HA under normoxic vs. hypoxic conditions (n = 5). ***P = 0.0001 in (D) refers to CD44 kd normoxia vs. Luc normoxia; **P = 0.002 in (D) refers to CD44 kd hypoxia vs. Luc hypoxia (E) EC tube formation over 4 h on growth factor-reduced Matrigel in the presence of CM harvested from 3D control vs. CD44 kd HT-29 spheres grown in Matrigel contain- ing HA under normoxic vs. hypoxic conditions (n = 5), scale = 100 lm. *P = 0.05 in (E) refers to CD44 kd normoxia vs. Luc normoxia; **P = 0.0001 in (E) refers to CD44 kd hypoxia vs. CD44 kd normoxia. Error bars represent standard deviation. Unpaired Student’s t-test was used in C, D, E.

Article Snippet: In addition, the corresponding cell culture supernatants (conditioned media, CM) of each well were harvested and analyzed using a VEGF ELISA kit (R&D systems, #DY293B) according to the manufacturer’s instructions.

Techniques: Biomarker Discovery, Immunostaining, Expressing, Cell Culture, Control, Standard Deviation