human vegf c quantikine elisa kit r d systems  (R&D Systems)

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: CRIP1 Reshapes the Gastric Cancer Microenvironment to Facilitate Development of Lymphatic Metastasis.

doi: 10.1002/advs.202303246

Figure Lengend Snippet: Figure 4. VEGFC is upregulated by CRIP1 in GC cells and involved in CRIP1-mediated lymphangiogenesis and LM. A,B) ELISA showing the effect of CRIP1 overexpression on VEGFC A) and VEGFD B) secretion. C,D) ELISA showing the effect of CRIP1 knockdown on VEGFC C) and VEGFD D) secretion. E,F) RT-qPCR showing the effect of CRIP1 overexpression E) and knockdown F) on VEGFC mRNA expression. G) Western blot showing the effect of CRIP1 overexpression and knockdown on VEGFC protein expression. H) Western blot showing the expression of CRIP1 and VEGFC in human GC samples. NT, nontumorous tissues; GC, gastric cancer tissues. I) Representative images (left panel) and quantification (right panel) of the Matrigel tube formation assay with human lymphatic endothelial cells (HLECs). HLECs were cultured with conditioned medium derived from GC cells treated as indicated. Scale bars = 200 μm. J) Representative images of enucleated popliteal lymph nodes across groups. K) Histogram analysis of lymph node volumes across groups. L) Ratios of metastatic to total dissected popliteal lymph nodes from mice inoculated with the indicated cells. 𝜒-square test was performed to assess the statistical significance in (L). Error bars represent the mean±SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: ELISA: Cell supernatants were collected to quantify the secretion of VEGFC (DVEC00, R&D systems, MN), VEGFD (DVED00, R&D systems, MN), CCL5 (mlbio, Shanghai, China), and TNF-α (mlbio, Shanghai, China) via ELISA assay.

Techniques: Enzyme-linked Immunosorbent Assay, Over Expression, Knockdown, Quantitative RT-PCR, Expressing, Western Blot, Tube Formation Assay, Cell Culture, Derivative Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: CRIP1 Reshapes the Gastric Cancer Microenvironment to Facilitate Development of Lymphatic Metastasis.

doi: 10.1002/advs.202303246

Figure Lengend Snippet: Figure 6. CREB1 mediates the CRIP1-regulated promotion of VEGFC expression and functions as a transcription factor for VEGFC. A,B) RT-qPCR showing the effect of CREB1 overexpression A) and knockdown B) on VEGFC mRNA expression. C,D) ELISA showing the effect of overexpression C) and knockdown D) of CREB1 on VEGFC secretion. E,F) Western blot analysis of p-CREB1 (Ser 133) and VEGFC expression in CRIP1 overexpression GC cells rescued by CREB1 knockdown E) and in CRIP1 knockdown GC cells rescued by CREB1 overexpression F). G,H) ELISA showing the expression of VEGFC in CRIP1 overexpression GC cells rescued by CREB1 knockdown G) and in CRIP1 knockdown GC cells rescued by CREB1 overexpression H). I) Representative images of enucleated popliteal lymph nodes for CREB1 overexpression cells (left panel). Histogram analysis of the lymph node volumes for groups (right panel). J) Representative images of enucleated popliteal lymph nodes for CREB1 knockdown cells (left panel). Histogram analysis of the lymph node volumes for groups (right panel). K) Specific primers were designed for potential CREB1 binding sites in the VEGFC promoter. L) Chromatin immunoprecipitation (ChIP) showing DNA fragments from the VEGFC promoter enriched in the DNA-protein complex immunoprecipitated via CREB1 antibody. M) Promoter luciferase assay showing that binding of CREB1 to the VEGFC promoter sites could enhance VEGFC transcription. Error bars represent the mean±SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: ELISA: Cell supernatants were collected to quantify the secretion of VEGFC (DVEC00, R&D systems, MN), VEGFD (DVED00, R&D systems, MN), CCL5 (mlbio, Shanghai, China), and TNF-α (mlbio, Shanghai, China) via ELISA assay.

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Knockdown, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Chromatin Immunoprecipitation, Immunoprecipitation, Luciferase