Journal: Mediators of Inflammation

Article Title: Leptin Induces an Inflammatory Phenotype in Lean Wistar Rats

doi: 10.1155/2009/738620

Figure Lengend Snippet: Effects of leptin on adipose tissue, serum adiponectin, and liver . Samples of mesenteric adipose tissue, serum and liver were collected from leptin-treated rats (1 mg/kg via the tail vein) and saline controls, and the following parameters were measured: Expressions of (a) MCP-1 and (b) IL-6 in samples of mesenteric adipose tissue were assessed using predeveloped assays for real-time PCR according to the manufacturer's instructions (Applied Biosystems). Values were calculated using a comparative C t method. (c) Serum levels of adiponectin were measured by ELISA. (d) Hepatic mRNA expression of ICAM-1 was measured by real-time PCR. Data are presented as mean ± SEM of at least 4 observations/group. Statistics were performed using Student's t -test * P < .05 compared to saline controls.

Article Snippet: DuoSet ELISA kits for Human ICAM-1, IL-6, and MCP-1 as well as ELISA kits for rat Adiponectin were purchased from R&D Systems (Minneapolis, MN).

Techniques: Saline, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Mediators of Inflammation

Article Title: Leptin Induces an Inflammatory Phenotype in Lean Wistar Rats

doi: 10.1155/2009/738620

Figure Lengend Snippet: Effect of leptin on hepatocytes . Human C3A hepatocytes were cultured in the presence of leptin (0–125 ng/mL) for 24 hours. (a) The release of soluble ICAM-1 into the culture medium and (b) ICAM-1 mRNA levels was quantified by ELISA and real-time PCR, respectively. Statistics were performed using one-way ANOVA * P < .05 compared to vehicle control cultures.

Article Snippet: DuoSet ELISA kits for Human ICAM-1, IL-6, and MCP-1 as well as ELISA kits for rat Adiponectin were purchased from R&D Systems (Minneapolis, MN).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Control

Journal: Mediators of Inflammation

Article Title: Leptin Induces an Inflammatory Phenotype in Lean Wistar Rats

doi: 10.1155/2009/738620

Figure Lengend Snippet: Effect of leptin on hepatocyte cytokine and chemokine release . Protein levels of IL-6 and MCP-1 produced by hepatocytes cultured in the presence of 62.5 ng/mL leptin for 24 hours were quantified using a high-sensitivity Quantikine ELISA kit (R&D systems, Inc). Data are presented as mean ± SEM of at least 4 observations/group. Statistical analysis was performed using Student's t -test * P < .05 compared to vehicle control cultures.

Article Snippet: DuoSet ELISA kits for Human ICAM-1, IL-6, and MCP-1 as well as ELISA kits for rat Adiponectin were purchased from R&D Systems (Minneapolis, MN).

Techniques: Produced, Cell Culture, Enzyme-linked Immunosorbent Assay, Control

Journal: Pharmaceuticals

Article Title: Pro-Apoptotic and Anti-EMT Activity of Wild Ginseng Adventitious Root Extract in MDA-MB-231 TNBC Cells: Association with GSK-3β/β-Catenin Signaling

doi: 10.3390/ph19020216

Figure Lengend Snippet: Proteome profiling of oncology-related markers in MDA-MB-231 cells treated with wild ginseng adventitious root extract (WGAR) and cisplatin. Cells were treated with WGAR or cisplatin at the 20% inhibitory concentration (IC20; 3 μg/mL for WGAR, 2 μg/mL for cisplatin) and 50% inhibitory concentration (IC50; 79 μg/mL for WGAR, 9 μg/mL for cisplatin) for 48 h. Cell lysates (200 μg) were analyzed using the Proteome Profiler Human XL Oncology Array (R&D Systems, ARY026). ( a ) Representative array membrane images. ( b ) Quantification of selected oncology-related markers at IC20 concentrations. Proteins are grouped by functional categories as indicated by horizontal lines below the x -axis. ( c ) Quantification of selected oncology-related markers at IC50 concentrations. Proteins are grouped by functional categories as indicated by horizontal lines below the x -axis. ( d ) Independent validation of IL-6 and M-CSF by ELISA. IL-6 and M-CSF levels in culture supernatants were quantified using the Human IL-6 Quantikine ELISA Kit (Cat# D6050, R&D Systems) and Human M-CSF Quantikine ELISA Kit (Cat# DMC00B, R&D Systems), respectively, and expressed as fold change relative to control (set to 1.0). Data are expressed as fold change relative to control (dashed line = 1). Values represent mean ± SD from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test: * p < 0.05, ** p < 0.01, *** p < 0.001 compared to the untreated control. CCL2, C-C motif chemokine ligand 2; CXCL8, C-X-C motif chemokine ligand 8; Dkk-1, Dickkopf-1; GM-CSF, granulocyte-macrophage colony-stimulating factor; IL-6, interleukin-6; M-CSF, macrophage colony-stimulating factor; PAI-1, plasminogen activator inhibitor-1; u-PA, urokinase-type plasminogen activator; VEGF, vascular endothelial growth factor; WGAR, wild ginseng adventitious root.

Article Snippet: To independently validate the proteome array findings, IL-6 and M-CSF levels in cell culture supernatants were quantified using the Human IL-6 Quantikine ELISA Kit (Cat# D6050, R&D Systems) and Human M-CSF Quantikine ELISA Kit (Cat# DMC00B, R&D Systems), respectively, according to the manufacturer’s instructions.

Techniques: Concentration Assay, Membrane, Functional Assay, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Control